Transporter systems, assemblies and associated methods for transporting tissue samples

ABSTRACT

A tissue sample that has been removed from a subject can be properly fixed for evaluation using the disclosed transporter assembly for carrying a tissue sample and method for fixing an unfixed tissue sample. In one embodiment, the disclosed assembly includes a transport container, a fixative in the transport container, and a cooling device that reduces and/or maintains the temperature of the fixative to perform a pre-soaking process at a temperature of less than about 7° C. The pre-soaking process can, for example, be performed during sample transport or during extended periods of storage, such as over a weekend.

CROSS-REFERENCE TO RELATED APPLICATIONS

This is a continuation of International Patent Application No.PCT/EP/2016/051489, filed Jan. 26, 2016, the benefit of which isclaimed. Benefit is further claimed to each of U.S. Provisional PatentApplication No. 62/107,874, filed Jan. 26, 2015, and U.S. ProvisionalPatent Application No. 62/108,184, filed Jan. 27, 2015. The contents ofall of these related applications are incorporated by reference herein.

FIELD

The present disclosure relates generally to transporter systems,assemblies and associated methods for transporting tissue samples. Morespecifically, the present disclosure relates to transporter systems andtransporters for monitoring and/or analyzing the condition of tissuesamples during transportation to improve the preservation of cellularactivity.

BACKGROUND

Preservation of tissues removed by surgical procedures is a topic ofgreat importance. After removal of a tissue sample from a subject, thetissue sample is often placed in a liquid that will suspend themetabolic activities of the cells. This process is commonly referred toas “fixation” and can be accomplished by several different types ofliquids. The most common fixative in use by anatomical pathology labs is10% neutral buffered formalin (NBF). This fixative forms cross-linksbetween formaldehyde molecules and amine containing cellular molecules.In addition, this type of fixative preserves proteins for storage. Whenused at room temperature, NBF diffuses into a tissue section andcross-links proteins and nucleic acids, thereby halting metabolism andpreserving the tissue for paraffin wax infiltration. The formalin can beat slightly elevated temperature (i.e., higher than room temperature) tofurther increase the cross-linking rate, whereas lower temperatureformalin can significantly decrease the cross-linking rate. For thisreason, histologists typically perform tissue fixation at roomtemperature or higher.

Several effects are often observed in tissues fixed by the abovemethods. First as the fixative is defusing into the tissue at room orhigher temperatures the cross-linking is occurring at the same time.This cross-linking inhibits the fixative diffusion which can allowenzymes in the tissue to degrade fragile biomarkers that can be criticalto disease interpretation. This also can lead to tissue that is eitherunder exposed or over exposed by liquid fixatives. For example, ifformalin has not diffused properly through the tissue samples, outerregions of the tissue samples exposed to formalin may be over-fixed andinterior regions of the tissue samples not exposed to formalin may beunder-fixed, resulting in very poor tissue morphology. In under-fixedtissue, subsequent exposure to ethanol often shrinks the cellularstructures and condenses nuclei since the tissues will not have thechance to form a proper cross-linked lattice. When under-fixed tissue isstained, such as with hematoxylin and eosin (H&E), many white spaces maybe observed between the cells and tissue structures, nuclei may becondensed, and samples may appear pink and unbalanced with thehematoxylin stain. Tissues that have been over-fixed typically do notwork well for subsequent immunohistochemical (IHC) processes. As aresult, the optimal antigen retrieval conditions for these tissues donot work properly and therefore the tissue samples appear to be understained.

Proper medical diagnosis and patient safety often require properlyfixing the tissue samples prior to staining. After excision, tissuesamples are often placed in warm formalin and transported from anoperating room to a laboratory. Unfortunately, the tissue samples tendto exhibit over-fixed outer regions and under-fixed inner regions. Itmay be difficult to identify such over-fixation and under-fixation uponarrival to the laboratory, thereby making it difficult to properlyprepare the samples for examination. The workers at the laboratory maynot know when to remove the tissue samples from the formalin or otherfixative. If tissue samples are removed from the formalin prior tocomplete diffusion and/or cross-linking, the sample may be under-fixed.If the specimen is left in the formalin an excess length of time,diagnosis may be delayed and the specimen may be over-fixed. Thisunder-fixation and over-fixation often results in poor preservation ofbiological molecules, tissue morphology, and/or post-translationalmodification signals.

Conventional, prefilled specimen containers are available from varioussuppliers. Such conventional containers offer only a simple lid forclosing after sample insertion. It is not possible to ascertain whatconditions the tissue sample in the container was exposed to or for howlong. Conventional specimen tracking is generally performed by ahandwritten process on paper labels or reports filled out after a samplehas been removed from a patient and before transporting the samplecontainer to the lab. Several minutes or even tens of minutes can lapsduring which sample degradation can occur.

There are several problems with current best-practice histology sample,collection, and transport methodologies. Current room-temperaturefixation protocols do not preserve biomarkers sufficiently. Tissuestorage after removal is not sufficiently standardized with respect tobiomarker preservation; this can result in some specimens even beingleft without formalin over the weekend in a refrigerator. The lack ofrigid adherence to set protocols across institutions and sites oftenresults in widely varying degrees of histomolecular degradation. Thiscompromises downstream processing, especially in samples subjected to avariety of biomarker-based diagnostics.

Current, standard specimen collection containers general contain asingle, initially-submerged tissue sample that does not stay in placeduring transport. These results in the tissue often floating up to thesurface, or adhered to the side of the wall of the container, thuscompromising the amount of contact each sample has with the fixativefluid, the perfusion of which is critical for proper preservation andsubsequent processing of the sample. Additionally, variations in ambienttemperature during transport are known to affect efficacy of fixative.WO 2012/040823 A1 discloses a two part specimen container where thefixative is held in a separate compartment until after the sample hasbeen placed in the specimen compartment. EP1913360B1 and U.S. Pat. No.8,691,165B2 ('360/'165 devices) disclose a specimen container that hastemperature and/or time data logging devices embedded in either thespecimen container, container cap or a tissue cassette carried in thespecimen container. None of the above systems/devices solve the fixationand other problems mentioned above.

General best-practices may vary intra- and inter-institutionally inspite of barcode labeling. Current methodologies rely heavily on paperand individualized computer input which may also vary from institutionto institution; chain of custody is established through a series ofsignatures. Real-time tracking of the computer specimen is paper-basedand cannot provide an up-to-the-minute accounting of temperature,location and transport time. The end result of these challenges isquestionable sample quality (biomarker preservation may not correctlyreflect the original state of the tissue) and lack of traceabilityduring processing. These are unacceptable risks when dealing withoncology patients.

SUMMARY OF THE DISCLOSURE

The disclosed sample collection, transport and tracking system includesan integrated solution for meeting one or more of the above-referencedchallenges, especially the preservation of biomarkers in a tissuesample. The various embodiments provide superior, digitized trackingthat begins the process of optimized formalin fixation and biomarkerpreservation without supervision by a laboratory technician. A cold partof the fixation process happens while the specimen is in transit. Thismeans that once the specimens reach their destination: an anatomicpathology laboratory, they need at most a short heat treatment (e.g. ˜1hour at 45° C.) to achieve substantially higher quality fixation forsubsequent processing, staining and analysis.

The disclosed integrated solution therefore can, in certain embodiments,result in 1) superior formalin fixation results for a broader range ofbiomarker-based diagnostics, including novel biomarker preservationpreviously only possible in fresh frozen specimens; 2) a processing timethat is compatible with standard fixation protocols (depending on tissuetype); 3) preservation of volatile biomarkers for up to 14 days prior tocomplete processing; 4) optionally, fully electronic trackingestablishing a chain of custody that can be readily accessed fromdigital tablets and/or uploaded for cloud computing; 5) optionally, areal-time recording of multiple conditions that can be used to gaugewhether additional measures will be necessary at destination to ensureoptimal processing, including staining.

In general, the presently disclosed sample collection, transport, andtracking systems can comprise: (a) a sample container for holding thetissue sample immersed in a volume of cold formalin-based fixativesolution; (b) a temperature-responsive element capable of tracking andrecording the temperature of formalin-based fixative solution disposedin the sample container; and (c) a case comprising an insulator or acooling element insulator and/or a cooling element, wherein the cavityis sized to accept the sample container and the temperature-responsiveelement in an arrangement that permits the temperature-responsiveelement to track and record the temperature of the formalin-basedfixative solution in the sample container, and wherein the insulatorand/or a cooling element is capable of maintaining the volume of theformalin-based fixative solution at a temperature in the range of 0° C.to 10° C. for at least 2 hours under shipping conditions when the tissuesample transport and storage assembly is assembled.

In some embodiments, an apparatus for processing tissue samples includesa container holding reagents, a carrier assembly, and a transportmechanism (transporter). The carrier assembly can include a specimenholder and a sensor or monitoring assembly positioned to monitor variousconditions of the tissue sample held by the specimen holder. In oneembodiment the monitor assembly can include one or more sensors selectedto, among other capabilities, determine, monitor or store data about:the temperature of the container and/or sample; elapsed time;environmental temperature in the transporter; ambient light to determineif the carrier assembly and sample container are in or outside thetransporter; acoustic; infrared proximity (determine if specimen or acassette containing the specimen is in the specimen container or thecontainer is in the carrier assembly, etc.); accelerometer to recordsimpact or inversion; leaks or fluid loss; or any other type of sensorsor sensor arrays. Additionally, one or more sensors can be configured totransmit data, such as temperature, for example, while being submergedin the reagent in the specimen holder in the reagents.

At least some embodiments disclosed herein are transporter systems formonitoring tissue samples before, during, and/or after fixation. Thetissue sample(s) are placed in fixative contained in a specimencontainer which is held in a carrier assembly which in turn is placed ina transporter. The specimen container can be removed from the carrierand transporter and processed based on information from a monitoringsystem. The monitoring system can provide, without limitation, diffusionor saturation information, fixation information, temperature-timeinformation, or other information about the tissue samples. In someprocedures, a transporter with a cooling capability can rapidly reducethe temperature of the fixative to a preferred temperature forminimizing or limiting fixation (e.g., cross-linking). The coolingtransporter can continuously or periodically cool the fixative to keepthe fixative at or below the pre-soaking temperature throughoutshipping. After fixative has adequately saturated the tissue sample, afixation process can later be performed to fix substantially the entiretissue sample. In some embodiments, the transporter can provideinformation to the laboratory indicating the diffusion or saturationstatus. In some embodiments, the cooling transporter is operable toreduce a temperature of the fixative to inhibit fixation of a tissuesample during a diffusion process.

At least some embodiments disclosed herein are specimen transportersthat can pre-cooled a fixative. A tissue sample can be contacted withthe pre-cooled fixative, which diffuses through the tissue sample. Thespecimen transporter can keep the tissue sample, and fixative diffusedthrough the tissue sample, at a relative low temperature to inhibit orprevent fixation. The fixation process can be caused by heating thetissue sample to start or promote cross-linking. In some embodiments,the transporter keeps the tissue sample at the low temperature duringtransport from a collection site (e.g., an operating room at a hospital)to a processing site (e.g., a pathology laboratory). After receiving thecontainer, the pathology laboratory can perform the fixation process. Assuch, substantially the entire fixation process can be performed at thepathology laboratory.

In some embodiments, a low temperature portable transporter systemincludes a carrier assembly including a holding chamber, a fixative in aspecimen container retained in the holding chamber, and either or both acooling device carried by a transporter or insulation in the transportersufficient to maintain the fixative at a desired temperature for adesired amount of time. The cooling device is generally in thermalcommunication with the specimen container and is operable to reduce atemperature of the fixative in the container. In one embodiment, thecarrier assembly has an open configuration for receiving the tissuesample and fixative container and a closed configuration for sealing theholding chamber. The cooling device can have a non-cooling mode and acooling mode. In one embodiment, the cooling device in the cooling modereduces a temperature of the fixative to at least about 15° C. in lessthan about 5 minutes. In some embodiments, the pre-soaking and laterfixation process preserve signals of proteins in the tissue samplesignificantly, for example, by preserving at least 30%, 50%, 70%, or 90%post-translation modification signals. The tissue fixation methods inaccordance with at least some embodiments can significantly halt theenzyme activities degrading proteins, such as halting the enzymeactivities of protease.

In some embodiments, the transporter system is adapted to perform anautonomous warming process, whereby a cold environment for the tissuesample container for a minimum period of time (generally at least 2hours) to allow for diffusion of the fixative solution into the tissues,before allowing it to passively warm to ambient temperature over a fewhours, at which temperature the tissue can be fixed through chemicalcrosslinking. The cold temperature is maintained by contact with athermal reservoir made of a phase change material which has a meltingtemperature close to the desired diffusion temperature (which isgenerally from 0 to 7° C.), which also allows the phase change materialto be frozen in conventional refrigerator. Exemplary phase changematerials include, for example, proprietary blends of oily (at roomtemperature) liquids, such as mixtures derived from vegetable oils(triglycerides), cycloalkanes and other components, and from paraffin(linear and/or branched hydrocarbons of various lengths).

In some embodiments, a portable transporter system includes a carrierassembly, a fixative, and a fixation-inhibiting cooling device. Thecarrier assembly includes a holding chamber containing the fixative. Thefixation-inhibiting cooling device is carried by the carrier assemblyand is in thermal communication with the fixative. Thefixation-inhibiting cooling device is operable to reduce a temperatureof the fixative to inhibit fixation of a tissue sample contacting thefixative. The portable transporter system can be readily carried (e.g.,manually transported) by a person. In a laboratory setting, the portabletransporter system can be manually transported between workstations andequipment. In a shipping setting, the portable transporter system can bemanually transported to and from a vehicle. The portable transporter canbe configured to thermally isolate or substance a pre-cool fixative.Additionally or alternatively, the portable transporter can beself-cooling to chill the fixative.

In yet further embodiments, a portable transporter system includes acarrier assembly, a fixative, and a thermal device operable to adjust ofmaintain the temperature of the fixative to inhibit fixation of a tissuesample, promote fixation, or the otherwise thermally process the tissuesample. The portable transporter system can be a handheld system forconvenient transport. In some embodiments, the thermal device includes afixation-inhibiting cooling device configured to reduce a temperature ofthe fixative to perform a diffusion process. In other embodiments, thethermal device includes a thermoelectric unit and/or refrigeration unit.

In one embodiment, a portable transporter system includes a carrierassembly holding fixative and fixation-inhibiting cooling means. Thecarrier assembly includes a holding chamber containing the fixative andsample. The fixation-inhibiting cooling means is in thermalcommunication with the fixative/sample container and is operable toreduce a temperature of the fixative from room temperature (e.g., about20° C. to about 25° C.) to a temperature (e.g., temperature less thanabout 15° C.). The fixation-inhibiting cooling means can include anactivation agent, a cooling agent, a refrigeration unit, or athermoelectric cooler unit. The portable transporter system can cool thefixative to the desired temperature. Alternatively, the portabletransporter system can keep a chilled, pre-cooled liquid at a desiredtemperature. Thus, the portable transporter system can reduce thetemperature of fixative to the pre-soak temperature or maintain thetemperature of the fixative already at the pre-soak temperature.

In some embodiments, an apparatus for processing tissue samples includesa container holding reagents, a carrier assembly, and a transportmechanism. The carrier assembly can include a specimen holder and asensor assembly positioned to determine the state of the tissue sampleheld by the specimen holder. The transport mechanism can submerge thesensor assembly and the tissue sample held by the specimen holder in thereagents. In one embodiment, the sensor is a temperature measurementsensor. In one embodiment, the apparatus further includes a computingdevice with instructions, when executed, that cause the apparatus to (a)if diffusion of fixative through the tissue sample at or above a targetdiffusion level, a fixation process is performed on the tissue sampleand (b) if diffusion of fixative through the tissue sample is below atarget diffusion level, a diffusion process is performed on the tissuesample. The target diffusion level can be selected based on the tissueprotocols to be performed. For example, in some protocols, the fixativecan be diffused throughout most of the volume of the tissue sample priorto fixation.

In one embodiment, the portable transporter system can further include acoupler for coupling the data collection device to the carrier assemblytogether to form a paired tissue sample with data collection.

In one embodiment, the portable transporter system can include anelectronic data logger operable to store data for at least one of:sensing presence or insertion of the tissue sample in the specimencontainer; the time of insertion of the tissue sample data; time ofinsertion of the tissue sample data in at least one memory; log time anddate data of at least one of: insertion in the specimen container of thetissue sample in the standard histology cassette, or receipt of a starttime data; start a timer to track time data from at least one of: aninsertion time of a the specimen container with the tissue sampletherein into the carrier assembly, or receipt of a start timer datasignal; determine a time duration data of at least one of: from the logtime and date, or from the start of the timer; indicate a time durationdata since at least one of: the log time and date, or the start of thetimer; indicate data via a display; indicate data via a liquid crystaldisplay (LCD); indicate data via a light emitting diode (LED) display;record a temperature data; record a temperature and time data; record atleast one of a temperature or a time of crossing at least onetemperature threshold data; record at least one of a temperature or timeof temperature excursion data against at least one preprogrammedthreshold; record at least one of a temperature or time of temperatureexcursion against heating or freezing data; record at least onetemperature change and a time of the at least one temperature changedata; indicate at least one recording; indicate on at least one displaythe at least one record; monitor temperature excursions data; monitor atleast one temperature change, and a time of the at least one temperaturechange, and log the at least one temperature change and the time data;monitor data of at least one temperature change and time after seal ofthe carrier assembly, and indicate relative performance of the dataagainst at least one pre-programmed threshold; indicate on at least onedisplay results data of the monitor; store data on at least one memory;store at least one of sensed data; recorded data; monitored data; orcalculated data on at least one memory; store date on at least onememory, wherein the data can include at least one of: a sampleidentifier; an identifier; a radio frequency identifier (RFID); a barcode identifier; QR code identifier; a location; a sensed location; areceived location; a global positioning system (GPS) location; alocation relative to a Wi-Fi access point; a location relative to awireless communication network; a wired communication data; a wirelesscommunication data; a direct connection; a local connection; a localarea network (LAN); a wide area network (WAN); a remote connection; aBluetooth network data; a near field communication (NFC) data; a Zigbeeprotocol-compliant standard wireless communication data; a date and timeof a data point; a sensed data point; a port connection; a universalserial bus (USB) port connection; a coupling to a communication device;a coupling to a computing device; a coupling to a portable device; acoupling to a wireless device; a coupling to a personal computer device;a coupling to a smartphone device; a coupling to a tablet device; acoupling to a mobile phone device; or a coupling to a telephony device.

In one embodiment, a timer apparatus is provided which uses diffusion ofa colored liquid through an absorbent membrane as a timer to indicatehow long the colored liquid has been in contact with the absorbentmembrane. Measurements of the time of arrival of the colored liquidfront at various points along the length of the absorbent membrane canbe used as a rough measurement of the time that a tissue sample has beenin fixative. In some embodiments, the timer apparatus is capable ofdetermining the amount of time that the fixative has spent at aparticular temperature range. For example, a device for monitoring thetime that a tissue sample has spent in room temperature fixative cancomprise an absorbent membrane separated from a dye-soaked pad by adye-impervious film. Once the film is removed, the dye comes intocontact with the absorbent membrane and starts to diffuse along itslength. The distance that the dye is seen to have diffused down themembrane indicates the length of time since the film was removed. Textand markings on the front of the device can either show the user howlong the tissue has been in fixative, or give an indication of the stateof the tissue. A similar device can measure the time that a tissuesample has spent at cold (e.g. 0-7° C.) and room temperature. A firstcolored dye indicates the length of time spent at cold temperature bythe same method of diffusion along an absorbent membrane and isinitiated by removal of the dye-impervious barrier. When the temperatureincreases beyond the desired temperature range, the dye should stopdiffusing and thus give a record of how long the tissue sample was at0-7° C. A second colored dye, now mixed with a material which melts at aspecific temperature above the cold temperature range (e.g. roomtemperature), will only diffuse along a separate absorbent membrane whenthe environment around the tissue sample reaches the specifictemperature. Thus one can also monitor the time that the tissue samplewas at or above the specific temperature. Since two times are beingmonitored, there are several possible states for the device to end in,indicating whether the tissue sample was exposed to both coldtemperature and a higher temperature for the proper amount of time forideal fixation. It is also possible for the device to merely report thelength of time (in hours) at each temperature. The device for monitoringthe time spent in cold and room temperature can include a method to haltthe diffusion of the first dye through the absorbent membrane when thetemperature increases in order to provide a record of how long thetissue sample spent in the cold temperature. For example, a phase changematerial is introduced behind the absorbent membrane in contact with thefirst dye. This phase change material (PCM) will remain solid at coldtemperature, but when the ambient temperature increases it will melt.The chemical composition of the PCM, absorbent membrane and first dyeshould be chosen such that the PCM fouls the absorbent membrane andprevents further diffusion of the first dye. For instance, the PCM couldbe a paraffin which melts at 18° C., and the first dye could be amixture of dye, water and glycerol. In that case the hydrophobicity ofthe paraffin absorbed into the membrane would prevent the aqueousmixture from diffusing further along the absorbent membrane.

In one embodiment, the portable transporter system can include afixative can prefilled into the specimen container, and can berefrigerated until ready for use.

In one embodiment, the tissue sample, when surgically removed, is markedwith a patient identifier.

In one embodiment, the portable transporter system where the patientidentifier can include at least one of: a barcode; a wristband; or aradio frequency identifier (RFID).

In one embodiment, the tissue sample is inserted into the standardhistology cassette and the standard histology cassette is placed in thecarrier assembly, wherein the standard histology cassette has a uniqueidentifier.

In one embodiment, the portable transporter system can include a carrierassembly, wherein the assembly holds a data logger in close proximity,and wherein the data logger logs at least one start time and starttemperature reading.

In one embodiment, the data logger logs temperatures continuously duringtransport to a laboratory.

In one embodiment, the portable transporter system can include, wherethe data logger logs at least one of a temperature or a time at leastone of: continuously; periodically; aperiodically; at least onethreshold; at least one temperature threshold; at a time interval; upona temperature change; according to sensed data; or after a timeduration.

In one embodiment, the portable transporter system where upon arrival atthe laboratory, time and temperature data is read from the data loggerof the transport assembly and verifies integrity of the tissue sample.

A tissue collection system is set forth to preserve volatilephosphomarkers and allow storage/transport for multiple days inconjunction with comprehensive chain-of-custody tracking. An exemplaryembodiment can include various exemplary components: a) a spill free andfume reduced handling and insertion of a specimen into an example samplecollection container for use with cold or room temperature (RT)fixatives (e.g. formalin of varying concentrations). Additionally aprovision to fully submerse the specimen in a fixative, to hold thesample specimen submersed, and to retain fluid contact even in aninverted orientation during transport. Additionally an option to use astandard tissue cassette to receive the sample and hold the cassettesuch that it is fully submersed in fixative during transport. Thecassette is presented in a dry and non-contaminating fashion for sampleinsertion. b) a comprehensive chain-of-custody tracking IT of histologyspecimen from point of removal in the operating room (OR) to HistologyLab. c) Embodiments provide for a Custom Tissue Transport Container withreduced exposures to fumes and risk of spills, as well as a coldtransport system with data monitoring. d) A method of comprehensivelylabeling and tracking specimen-ID from the OR to microscope slide usingidentification schemes such as, e.g., but not limited to, RFID tags,magnetic strips, QR codes, ePens, and barcodes, etc. connected by mobile(and/or static) computer systems is set forth. An improved comprehensivemethod of using one consistent specimen ID methodology between PatientID to embedded tissue block, to ID used on microscope slide used forstaining, and to imaging, diagnosis, and final report on computer is setforth.

At least some embodiments disclosed herein are transporter systemsconfigured to monitor tissue samples before, during, and/or afterfixation. The tissue samples are pre-soaked in fixative contained in atransporter. The tissue samples can be removed from the transporter andprocessed based on information from a monitoring system. The monitoringsystem can provide, without limitation, diffusion or saturationinformation, fixation information, temperature-time information, orother information about the tissue samples. In some procedures, aself-cooling transporter can rapidly reduce the temperature of thefixative to a pre-soaking temperature thereby minimizing or limitingfixation (e.g., cross-linking). The self-cooling transporter cancontinuously or periodically cool the fixative to keep the fixative ator below the pre-soaking temperature throughout shipping. After fixativehas adequately saturated the tissue sample, a fixation process can beperformed to fix substantially the entire tissue sample. In someembodiments, the transporter can provide information to the laboratoryindicating the diffusion or saturation status. In some embodiments, theself-cooling transporter is operable to reduce a temperature of thefixative to inhibit fixation of a tissue sample during a diffusionprocess.

At least some embodiments disclosed herein are specimen transportersthat can pre-cool a fixative. A tissue sample can be contacted with thepre-cooled fixative, which diffuses through the tissue sample. Thespecimen transporter can keep the tissue sample, and fixative diffusedthrough the tissue sample, at a relative low temperature to inhibit orprevent fixation. The fixation process can be caused by heating thetissue sample to start or promote cross-linking. In some embodiments,the transporter keeps the tissue sample at the low temperature duringtransport from a collection site (e.g., an operating room at a hospital)to a processing site (e.g., a pathology laboratory). After receiving thecontainer, the pathology laboratory can perform the fixation process. Assuch, substantially the entire fixation process can be performed at thepathology laboratory.

In some embodiments, a self-cooling transporter system includes atransport container including a holding chamber, a fixative in theholding chamber, and a cooling device carried by the transportcontainer. The cooling device is in thermal communication with thefixative and is operable to reduce a temperature of the fixative. In oneembodiment, the transport container has an open configuration operativeto receive the tissue sample and a closed configuration operative toseal the holding chamber. The cooling device has a non-cooling mode anda cooling mode. In one embodiment, the cooling device in the coolingmode reduces a temperature of the fixative at least about 25° C. in lessthan about 5 minutes. In some embodiments, the pre-soaking and fixationprocess preserve signals of proteins in the tissue sample significantly,for example, by preserving at least 30%, 50%, 70%, or 90%post-translation modification signals. The tissue fixation methods inaccordance with at least some embodiments can significantly halt theenzyme activities degrading proteins, such as halting the enzymeactivities of protease.

In some embodiments, a portable transporter system includes a transportcontainer, a fixative, and a fixation-inhibiting cooling device. Thetransport container includes a holding chamber containing the fixative.The fixation-inhibiting cooling device is carried by the transportcontainer and is in thermal communication with the fixative. Thefixation-inhibiting cooling device is operable to reduce a temperatureof the fixative to inhibit fixation of a tissue sample contacting thefixative. The portable transporter system can be readily carried (e.g.,manually transported) by a person. In a laboratory setting, the portabletransporter system can be manually transported between workstations andequipment. In a shipping setting, the portable transporter system can bemanually transported to and from a vehicle. The portable transporter canbe conFigured to thermally isolate or substance a pre-cool fixative.Additionally or alternatively, the portable transporter can beself-cooling to chill the fixative.

In yet further embodiments, a portable transporter system includes atransport container, a fixative, and a thermal device operable to adjustthe temperature of the fixative to inhibit fixation of a tissue sample,promote fixation, or the otherwise thermally process the tissue sample.The portable transporter system can be a handheld system for convenienttransport. In some embodiments, the thermal device includes afixation-inhibiting cooling device configured to reduce a temperature ofthe fixative to perform a diffusion process. In other embodiments, thethermal device includes a thermoelectric unit and/or refrigeration unit.

In one embodiment, a self-cooling portable transporter system includes atransport container holding fixative and fixation-inhibiting coolingmeans. The transport container includes a holding chamber containing thefixative. The fixation-inhibiting cooling means is in thermalcommunication with the fixative and is operable to reduce a temperatureof the fixative from room temperature (e.g., about 20° C.-about 25° C.)to a pre-soak temperature (e.g., temperature less than about 5° C.). Thefixation-inhibiting cooling means can include an activation agent, acooling agent, a refrigeration unit, or a thermoelectric cooler unit.The portable transporter system can pre-cool the fixative.Alternatively, the portable transporter system can keep a chilled liquidat a desired temperature. Thus, the portable transporter system canreduce the temperature of fixative to the pre-soak temperature ormaintain the temperature of the fixature already at the pre-soaktemperature.

In some embodiments, an apparatus for processing tissue samples includesa container holding reagents, a carrier assembly, and a transportmechanism. The carrier assembly can include a specimen holder and asensor assembly positioned to acoustically analyze the tissue sampleheld by the specimen holder. The transport mechanism can submerge thesensor assembly and the tissue sample held by the specimen holder in thereagents. In one embodiment, the apparatus further includes a computingdevice with instructions, when executed, that cause the apparatus to (a)if diffusion of fixative through the tissue sample at or above a targetdiffusion level, a fixation process is performed on the tissue sampleand (b) if diffusion of fixative through the tissue sample is below atarget diffusion level, a diffusion process is performed on the tissuesample. The target diffusion level can be selected based on the tissueprotocols to be performed. For example, in some protocols, the fixativecan be diffused throughout most of the volume of the tissue sample priorto fixation.

In one embodiment, a portable transporter system adapted to carry atissue sample in a standard histology cassette, the tissue samplecontacting a fixative, can include: a transport container can include: aholding chamber; a cassette septum; and a fixative in the holdingchamber; and a container lid can include: a cassette clip attachablycouplable to receive, hold and retain the standard histology cassette.

In an embodiment, the portable transporter system can further include:fixation-inhibiting cooling device carried by the transport containerand in thermal communication with the fixative in the holding chamber,wherein the fixation-inhibiting cooling device is operable to reduce atemperature of the fixative to inhibit fixation of the tissue samplecontacting the fixative.

In one embodiment, the portable transporter system can further include acassette cover removably coupled to the container lid.

In one embodiment, the portable transporter system can include where thecassette cover can include a sterile seal between the cassette cover andthe container lid.

In one embodiment, the portable transporter system can include where thefixative can include formalin.

In one embodiment, the portable transporter system can include where thecassette septum is removable with the container lid, the cassette clip,and the standard histology cassette.

In one embodiment, the portable transporter system can include where theremovable cassette septum can provide easy access to the standardhistology cassette while eliminating splashing of the fixative.

In one embodiment, the portable transporter system can include where thecassette clip is removable couplable to the container lid.

In one embodiment, the portable transporter system can include where thecontainer clip is operable to be screwed into at least one side of thecontainer lid.

In one embodiment, the portable transporter system where the cassetteclip is operable to be clipped to the standard histology cassette.

In one embodiment, the portable transporter system can include where thecassette clip includes: a first end operable to be coupled to at leastone side of the container lid; and a second end operable to be coupledto the standard histology cassette.

In one embodiment, the portable transporter system can include where thecassette septum can include a breakable membrane.

In one embodiment, the portable transporter system can include where thecontainer lid can further include an arm operable to break the breakablemembrane upon insertion of the cassette into the holding chamber of thetransport container.

In one embodiment, the portable transporter system can include where thecontainer lid can further include a cylindrical portion, which caninclude an internal screw thread portion adapted to couple with anexternal screw thread portion of the transport container, operable toisolate the sample, air and the fixative from a surrounding environment.

In one embodiment, the portable transporter system can further include adata collection device.

In one embodiment, the portable transporter system can further include acoupler for coupling the data collection device to the transportcontainer together to form a paired tissue sample with data collection.

In one embodiment, the portable transporter system can further include afirst shipping receptacle portion for receiving a plurality of thepaired tissue samples with data collections.

In one embodiment, the portable transporter system can further include asecond shipping receptacle portion for receiving the plurality of thepaired tissue samples with data collections.

In one embodiment, the portable transporter system can further includeat least one fixation-inhibiting cooling device in thermal communicationwith the fixative in the holding chamber, wherein thefixation-inhibiting cooling device is operable to reduce a temperatureof the fixative to inhibit fixation of the tissue sample contacting thefixative.

In one embodiment, the portable transporter system can further includeat least one cold transport box operable to receive the first shippingreceptacle portion, the plurality of the paired tissue samples with datacollections, and the at least one fixation-inhibiting cooling device.

In one embodiment, the portable transporter system wherein the cassetteseptum can include at least one split.

In one embodiment, the portable transporter system can include where theat least one split of the cassette septum includes at least two splits.

In one embodiment, the portable transporter system where the at leasttwo splits of the cassette septum can include where the at least twosplits are perpendicular to one another.

In one embodiment, the portable transporter system can include where theat least two splits of the cassette septum can include wherein the atleast two splits are of different length to one another.

In one embodiment, the portable transporter system where the at leastone split of the cassette septum can include at least one cross-shapedsplit.

In one embodiment, the portable transporter system can include where thestandard histology cassette holding the tissue sample is operable tomaintain the tissue sample submersed in fluid independent of theorientation of the transport container.

In one embodiment, the portable transporter system where the cassetteclip can include a clip feature operable to hold and retain the standardhistology cassette, where the standard histology cassette is adapted toinclude a corresponding feature.

In one embodiment, the portable transporter system can further include acarrier feature operable to slideably receive and retain the standardhistology cassette therein.

In one embodiment, the portable transporter system can include where thecontainer lid can include at least one side operable to hold an unusedcassette upward to avoid contamination with the fixative prior to sampleinsertion.

In one embodiment, the portable transporter system can include where thecontainer lid can include at least one side operable to seal thefixative fluid inside the holding chamber of the transport containerduring transport or handling and to avoid spillage and fumes.

In one embodiment, the portable transporter system where the containerlid can include: at least one side operable to releasably couple thecontainer lid onto the holding chamber.

In one embodiment, the portable transporter system can include where theat least one side operable to releasably couple the container lid ontothe holding chamber couples via matching screw threads.

In one embodiment, the portable transporter system can include where thestandard histology cassette is held separately until the tissue sampleis collected, and where the container lid is operable to receive thestandard histology cassette when the tissue sample is ready forinsertion into the holding chamber.

In one embodiment, the portable transporter system can include where thecontainer lid can include: a multi-purpose lid wherein at least one sideis operable to at least one of: hold an unused cassette upward to avoidcontamination with the fixative prior to sample insertion; seal thefixative fluid inside the holding chamber of the transport containerduring transport or handling and to avoid spillage and fumes; releasablycouple the container lid onto the holding chamber; releasably couple thecontainer lid onto the holding chamber couples via matching screwthreads; or receive the cassette when the tissue sample is ready forinsertion.

In one embodiment, the portable transporter system, can further includean electronic data logger coupled to the transport container via a pod.

In one embodiment, the portable transporter system can include where theelectronic data logger is in thermal communication with the transportcontainer.

In one embodiment, the portable transporter system can include where theelectronic data logger is operable to at least one of: sense insertionof the tissue sample in the transport container; determine time ofinsertion of the tissue sample data; store time of insertion of thetissue sample data in at least one memory; log time and date data of atleast one of: insertion in the transport container of the tissue samplein the standard histology cassette, or receipt of a start time data;start a timer to track time data from at least one of: an insertion timeof the standard histology cassette with the tissue sample in thetransport container, or receipt of a start timer data signal; determinea time duration data of at least one of: from the log time and date, orfrom the start of the timer; indicate a time duration data since atleast one of: the log time and date, or the start of the timer; indicatedata via a display; indicate data via a liquid crystal display (LCD);indicate data via a light emitting diode (LED) display; record atemperature data; record a temperature and time data; record at leastone of a temperature or a time of crossing at least one temperaturethreshold data; record at least one of a temperature or time oftemperature excursion data against at least one preprogrammed threshold;record at least one of a temperature or time of temperature excursionagainst heating or freezing data; record at least one temperature changeand a time of the at least one temperature change data; indicate atleast one recording; indicate on at least one display the at least onerecord; monitor temperature excursions data; monitor at least onetemperature change, and a time of the at least one temperature change,and log the at least one temperature change and the time data; monitordata of at least one temperature change and time after seal of thetransport container, and indicate relative performance of the dataagainst at least one pre-programmed threshold; indicate on at least onedisplay results data of the monitor; store data on at least one memory;store at least one of sensed data; recorded data; monitored data; orcalculated data on at least one memory; store date on at least onememory, wherein the data can include at least one of: a sampleidentifier; an identifier; a radio frequency identifier (RF ID); a barcode identifier; QR code identifier; a location; a sensed location; areceived location; a global positioning system (GPS) location; alocation relative to a Wi-Fi access point; a location relative to awireless communication network; a wired communication data; a wirelesscommunication data; a direct connection; a local connection; a localarea network(LAN); a wide area network (WAN); a remote connection; aBluetooth network data; a near field communication (NFC) data; a Zigbeeprotocol-compliant standard wireless communication data; a date and timeof a data point; a sensed data point; a port connection; a universalserial bus (USB) port connection; a coupling to a communication device;a coupling to a computing device; a coupling to a portable device; acoupling to a wireless device; a coupling to a personal computer device;a coupling to a smartphone device; a coupling to a tablet device; acoupling to a mobile phone device; or a coupling to a telephony device.

In one embodiment, the portable transporter system can further include acarrier unit integrated with a plurality of the container lids.

In one embodiment, the portable transporter system can include where theintegrated carrier unit can include where the plurality of the containerlids can include at least four container lids.

In one embodiment, the portable transporter system can further include aprotective sheath operable to protect the standard histology cassetteagainst contamination or dust.

In one embodiment, the portable transporter system where the carrierunit can include a plurality of holding chambers shipped with fixativein the plurality of holding chambers, each having one of the containerlids.

In one embodiment, the portable transporter system can include where thecontainer clip does not require the standard histology cassette to beremoved from the container lid.

In one embodiment, the portable transporter system can include where thecontainer lid is reversible and can be screwed onto the transportcontainer immersing the tissue sample in the fixative.

In one embodiment, the portable transporter system, where it is possibleat a glance to see which of the plurality of holding chambers havesamples inserted.

In one embodiment, the portable transporter system can include where thefixative can include formalin.

In one embodiment, the portable transporter system can include whereeach of the container lids is tall enough to receive application of atleast one label, code, barcode, or identifier.

In one embodiment, the portable transporter system can include whereeach the transport container cannot be inadvertently left outside of afixation-inhibiting cooling device.

In one embodiment, the portable transporter system can include wherecassette clip holds the standard histology cassette to ease reading ofat least one code on the standard histology cassette.

In one embodiment, the portable transporter system can include where thefixative can be prefilled into the transport container, and can berefrigerated to be ready for use.

In one embodiment, the portable transporter system can include where thetissue sample, when surgically removed is marked with a patientidentifier.

In one embodiment, the portable transporter system where the patientidentifier can include at least one of: a barcode; a wristband; or aradio frequency identifier (RFID).

In one embodiment, the portable transporter system can include where thetissue sample is inserted into the standard histology cassette, and thestandard histology cassette is placed in the transport container,wherein the standard histology cassette has a unique identifier.

In one embodiment, the portable transporter system can include where thetransport container is transferred into a pod, wherein the pod holds adata logger in close proximity, and wherein the data logger logs atleast one start time and start temperature reading.

In one embodiment, the portable transporter system can include where thepod is inserted into a fixation-inhibiting cooling device.

In one embodiment, the portable transporter system can include where thefixation-inhibiting cooling device can ensure the tissue sample is keptat 4° C. during transport.

In one embodiment, the portable transporter system can include where thedata logger logs temperatures continuously during transport to alaboratory.

In one embodiment, the portable transporter system can include, wherethe data logger logs at least one of a temperature or a time at leastone of: continuously; periodically; aperiodically; at at least onethreshold; at at least one temperature threshold; at a time interval;upon a temperature change; according to sensed data; or after a timeduration.

In one embodiment, the portable transporter system where upon arrival atthe laboratory, time and temperature data is read from the data loggerof the pod of the transport container and verifies integrity of thetissue sample.

In one embodiment, the portable transporter system can include where thetissue sample is stored in cold storage if additional cold storage timeis determined to be required.

In one embodiment, the portable transporter system can include, wherethe tissue sample is reviewed and at least one of transferred or kept ina final code identified standard histology cassette.

In one embodiment, the portable transporter system can include, wherethe tissue sample is processed using a standard heated fixativeprocessor, and with subsequent processing steps.

In one embodiment, the portable transporter system can include whereinthe standard heated fixative processor processing the tissue sample for2 hours at 45° C. temperature.

In one embodiment, the portable transporter system can include whereinthe subsequent processing steps can include at least one of an alcoholprocessing step, a xylene processing step, or a paraffin processingstep.

In one embodiment, the portable transporter system can include where thetissue sample is embedded, sectioned, and stained.

In one embodiment, the portable transporter system can include where thetissue sample is stained with various markers or probes.

In one embodiment, the portable transporter system can include where thestain results are made available for at least one of: review by apathologist, receipt of at least one score by a pathologist; or reportresults for the tissue sample to at least one clinical database.

In one embodiment, the portable transporter system can further includewhere electronic data is collected.

In one embodiment, the portable transporter system can include where theelectronic data can be collected by at least one of: an electronic pen;a unique identifier; a tablet device; a wireless identifier; a Wi-Fimedia access control (MAC) address; a static computing device; a mobilecomputing device; a barcode scanner; an external data temperaturelogger; a portable fixation-inhibiting cooling device can includeintelligence; a portable fixation-inhibiting cooling device can includean electronic datalogger for collecting temperature and time data;

In one embodiment, the portable transporter system can include where afixation-inhibiting cooling element is pre-charged over night for nextday surgery.

In one embodiment, the portable transporter system can include where thetransport container can be kept pre-cooled in a refrigerator.

In one embodiment, the portable transporter system can include where afixation-inhibiting cooling device is configured prior to surgery, istransported to surgery location, and keeps reagents cold.

In one embodiment, the portable transporter system can include wheretransportation of the portable transporter system is performed at an endof a surgery day.

In one embodiment, the portable transporter system can include where afixation processor is operable to at least one of: collect transportdata; perform final fixation steps required; or provide a comprehensivefixation quality report for each of the tissue samples.

In one embodiment, the portable transporter system can include where theelectronic data is collected by an electronic pen, and collection caninclude processing of data of the tissue sample can include at least oneof: establish chain-of-custody during the tissue sample collection,integrate data can include at least one of: patient-ID, location ofsurgery, day/time, name of surgeon, or location of removal of the tissuesample; associate with a designated of the transport container andcassette identifier (ID); record insertion time of the tissue sampleinto the transport container; log transport conditions can include atleast one temperature or time profile; document a total time of colddiffusion after which the sample can be safely removed and submitted toa heated fixative step for crosslinking; duplicate capture ofhandwritten Lab Requisition Form by electronic data capture andassociate data to a specific specimen of the tissue sample or thestandard histology cassette via identifier can include at least one of abarcode or radio-frequency identifier (RFID); verify a minimum fixationcondition; or enable data capture for a clinical trial.

In one embodiment, the portable transporter system can include where theelectronic data is collected by an electronic pen (ePen) can include atleast one of: use ePen to fill out lab requisition form; collectelectronic data; collect identifier information from patient, thetransport container, the standard histology cassette; transferelectronic data to a computing device for verification or correction;upload verified electronic data via communications network if possible;if upload not possible, ePen travels with the tissue sample infixation-inhibited cooling device; receive at accessioning stage inhistology lab for display and report to the LIS; optionally enter anycorrections, if required; or provide readout of datalogger, or integrateinto data uploaded to the LIS.

In one embodiment, the portable transporter system can include where theelectronic data is collected by a unique identifier.

In one embodiment, the portable transporter system can include, whereinthe unique identifier can include at least one of: an Anoto code printedonto a label of a container; a barcode on the cassette could be avantage compatible preprinted code being inserted into the container; abarcode on the cassette identifiable by a barcode reader when thecontainer arrives at accessioning stage, wherein the barcode reader istied to patient-ID associated to a code of the container; or a uniquelabel with Anoto pattern on the container, wherein the container ispicked randomly, pen ties code electronically to patient-ID, andhandwritten link and an identifier to be copied from a label of thecontainer onto paper.

In one embodiment, the portable transporter system can include where theelectronic data is collected by a tablet device coupled to the portabletransporter system by at least one of wired or wireless communication,and collection can include processing of data of the tissue sample caninclude at least one of: establish chain-of-custody during the tissuesample collection, integrate data can include at least one of:patient-ID, location of surgery, day/time, name of surgeon, or locationof removal of the tissue sample; associate with a designated of thetransport container and cassette identifier (ID); record insertion timeof the tissue sample into the transport container; log transportconditions can include at least one temperature or time profile;document a total time of cold diffusion after which the sample can besafely removed and submitted to a heated fixative step for crosslinking;duplicate capture of handwritten Lab Requisition Form by electronic datacapture and associate data to a specific specimen of the tissue sampleor the standard histology cassette via identifier can include at leastone of: a barcode; or radio-frequency identifier (RFID); data capturevia at least one of: camera; RFID; or near field communication (NFC) tagvia communication interface; capture of image of surgical site; verify aminimum fixation condition; or enable data capture for a clinical trial.

In one embodiment, the portable transporter system can include where theelectronic data is collected by the tablet can include at least one of:capture pertinent information about patient and study on the tablet;capture duplicate entry of lab requisition form on the tablet;optionally correct any data entry errors on screen, if needed;optionally upload verified electronic data via wireless communication toLIS; provide tablet accompanying the tissue sample in same cooling andshipping box; receive at the accessioning stage in histology lab thetablet for display or report to the LIS; or provide readout ofdatalogger, or integrate into data uploaded to the LIS.

In one embodiment, the portable transporter system can include whereinthe electronic data is collected by a wireless identifier.

In one embodiment, the portable transporter system can include where theelectronic data is collected by the wireless identifier can include atleast one of: a wireless fidelity (Wi-Fi) media access control (MAC)address.

In one embodiment, the portable transporter system can include where theelectronic data is collected by the Wi-Fi MAC address and wherein adevice having the Wi-Fi MAC address can include at least one of: adatalogger and positional time stamper operable to track the tissuesample; any time a Wi-Fi router or device with the MAC addresscommunicate, identity of both the router and the device are logged; orsince time of contact is known and location of a Wi-Fi router is known,a location of a device with the MAC address is recorded over the journeyof the MAC device via a network of free-access Wi-Fi routers.

In one embodiment, the portable transporter system can include whereinthe electronic data is collected by a static computing device, andcollection can include processing of data of the tissue sample caninclude at least one of: establish chain-of-custody during the tissuesample collection, integrate data can include at least one of:patient-ID, location of surgery, day/time, name of surgeon, or locationof removal of the tissue sample; associate with a designated of thetransport container and cassette identifier (ID); record insertion timeof the tissue sample into the transport container; log transportconditions can include at least one temperature or time profile;document a total time of cold diffusion after which the sample can besafely removed and submitted to a heated fixative step for crosslinking;duplicate capture of handwritten Lab Requisition Form by electronic datacapture and associate data to a specific specimen of the tissue sampleor the standard histology cassette via identifier can include at leastone of: a barcode; or radio-frequency identifier (RFID); data capturevia at least one of: camera; RFID; or near field communication (NFC) tagvia communication interface; capture of image of surgical site; verify aminimum fixation condition; or enable data capture for a clinical trial.

In one embodiment, the portable transporter system can include where theelectronic data is collected by the static computing device, can includeat least one of: capture pertinent information about patient and study;capture duplicate entry of lab requisition form; optionally correct anydata entry errors on screen, if needed; optionally upload verifiedelectronic data via communication network to LIS; if LIS connectivity isprohibited, optionally program data logger to retain additionalinformation on computer locally; receive at the accessioning stage inhistology lab identifier information to reference the tissue sample viaLIS query for display or report to the LIS; or provide readout ofdatalogger, or integrate into data uploaded to the LIS.

In one embodiment, the portable transporter system can include where theelectronic data is collected by a travelling cooling shipping box withintelligence, and collection can include processing of data of thetissue sample can include at least one of: establish chain-of-custodyduring the tissue sample collection, integrate data can include at leastone of: patient-ID, location of surgery, day/time, name of surgeon, orlocation of removal of the tissue sample; associate with a designated ofthe transport container and cassette identifier (ID); record insertiontime of the tissue sample into the transport container; log transportconditions can include at least one temperature or time profile;document a total time of cold diffusion after which the sample can besafely removed and submitted to a heated fixative step for crosslinking;duplicate capture of handwritten Lab Requisition Form by electronic datacapture and associate data to a specific specimen of the tissue sampleor the standard histology cassette via identifier can include at leastone of: a barcode; or radio-frequency identifier (RFID); data capturevia at least one of: camera; RFID; or near field communication (NFC) tagvia communication interface; capture of image of surgical site; verify aminimum fixation condition; or enable data capture for a clinical trial.

In one embodiment, the portable transporter system can include where theelectronic data is collected by the travelling cooling shipping box withthe intelligence, can include at least one of: capture pertinentinformation about patient and study; capture duplicate entry of labrequisition form; optionally interface computer with a host computer inthe travelling cooling shipping box to store data; optionally monitorloading or unloading of the tissue sample by the host computer in thetravelling cooling shipping box, and log time and temperature;optionally correct any data entry errors on screen, if needed; or at theaccessioning stage in histology lab, the host computer provideselectronic data collected.

In one embodiment, the portable transporter system can include where thetransport of the tissue sample can include a cooling box configured toat least one of: pre-charge over night for a next day surgery; pre-coolin fridge the container; prior to surgery transport to surgery locationand maintain in cold temperature reagents; or initiate shipping at endof surgery day.

In one embodiment, the portable transporter system can include where thecooling box is maintained at about 4° C.

In one embodiment, a portable transporter system adapted to carry atissue sample contacting a fixative, can include: a transport containercan include: a holding chamber; and a fixative; and a container lid caninclude: a cassette holder operable to be coupled to the standardhistology cassette and can include a first seal; and a cassette receiveroperable to be releasably coupled to the transport container and caninclude a second seal, where the cassette holder and the cassettereceiver are operable to be attachably coupled to one another uponinsertion of the standard histology cassette coupled to the cassetteholder into the cassette receiver.

In one embodiment, the portable transporter system can include, whereinthe cassette holder and the cassette receiver are operable to beattachably coupled to one another via at least one male snap clip and atleast one female snap clip receiver.

In one embodiment, the portable transporter system can include where theat least one male snap clip is on a bottom portion of the cassetteholder, and the at least one female snap clip receiver is on an upperportion of the cassette receiver.

In one embodiment, the portable transporter system can include where theat least one male snap clip can include a plurality of the at least onemale snap clips.

In one embodiment, the portable transporter system can include where theplurality of the male snap clips can include at least four snap clips.

In one embodiment, the portable transporter system can include where thefirst and second seals comprise solvent compatible o-rings.

In one embodiment, the portable transporter system can include where thefixative has a fixative volume can include: a holding chamber volume ofthe holding chamber less a cassette volume of a standard histologycassette.

In one embodiment, a portable transporter system adapted to carry atissue contacting a fixative, can include a transport container caninclude: a holding chamber; and a container lid operable to bereleasably coupled to the transport container, the container lid caninclude: a built-in septum can include a molded boss can include anexternal thread; and a standard sealing screw-top can include an innerthread matching the external thread of the molded boss, the screw-topoperable to cover the built-in septum.

In one embodiment, the portable transporter system can further includewhere a syringe can include a pre-filled amount of fixative, andoperable to be refrigerated until used to fill the holding chamber withthe fixative, via the built-in septum, after the tissue sample is placedinside the holding chamber.

In one embodiment, the portable transporter system can include where thefixative can include formalin.

In one embodiment, the portable transporter system can further includewhere the a cassette clip attachably couplable to receive, hold andretain the standard histology cassette.

BRIEF DESCRIPTION OF THE DRAWINGS

Non-limiting and non-exhaustive embodiments are described with referenceto the following drawings. The same reference numerals refer to likeparts or acts throughout the various views, unless otherwise specified.

FIG. 1 is a workflow for processing a tissue sample from a collectionsite to a processing site in accordance with one embodiment.

FIG. 2 is an exploded isometric view of a transport system in accordancewith one embodiment.

FIG. 3 is an illustration of an exemplary four cup transport system.

FIG. 4A is an exemplary carrier assembly including an exemplary TEMPODcasing with an exemplary specimen vial with specimen lid fixed atop it,and an exemplary data logger, both inserted in exemplary slots therein,according to an exemplary embodiment.

FIG. 4B is an example TEMPOD data logger.

FIG. 4C is an example CRYOPAK data logger usable for both cold- androom-temperature transport, according to another exemplary embodiment.

FIG. 5A illustrates an example initial paper-based form.

FIGS. 5B and 5C are various exemplary views of an electronic pen.

FIGS. 6A and 6B depict an exemplary exploded view, and inserted view,respectively, of an exemplary specimen vial, with specimen lid, cassettecoupled to lid by an exemplary clip and pod casing, and exemplaryCRYOPACK data logger according to an exemplary embodiment.

FIGS. 7A-7G are exemplary carrier assemblies.

FIGS. 8A-8H are exemplary carrier assembly data loggers.

FIGS. 9A and 9B show another exemplary carrier assembly data loggers.

FIG. 10A is an exemplary digital pen with exemplary functionality andcomponents.

FIG. 10B is another exemplary digital pen and components.

FIG. 11 is an exemplary process flow diagram illustrating an exampleworkflow variation between paper forms and e-LRF pen, illustrating formscreated and distributed by PharmaForms.

FIG. 12 is an exemplary process flow diagram illustrating an exampleworkflow variation between paper forms and e-LRF pen, illustrating formscreated by a responsible party.

FIG. 13 is an exemplary process flow diagram illustrating an exampleworkflow variation between paper forms and e-LRF pen, illustratingexample information flow from pen to laboratory information system(LIS).

FIG. 14 is an exemplary sample collection and tracking system (SCTS).

FIG. 15 is an example of tracking via an example electronic pen.

FIG. 16 is an example of tracking via an example electronic tabletdevice.

FIG. 17 is an example of tracking via an example static computer at thecollection site.

FIG. 18 is an example of tracking via an example travellingcooling/shipping box with intelligence.

FIG. 19 is an example of an example cooling box for specimen transport.

FIG. 20A is a workflow for processing a tissue sample in accordance withone embodiment.

FIG. 20B is a workflow for processing a tissue sample from a collectionsite to a processing site in accordance with one embodiment.

FIG. 20C is a front view of a self-cooling transporter system inaccordance with one embodiment.

FIG. 20D is a cutaway view of the transporter system of FIG. 20C.

FIG. 20E is a cross-sectional view of the transporter system of FIG. 2.

FIG. 20F is a cross-sectional view of specimen container in accordancewith one embodiment.

FIG. 20G is a perspective view of a transporter system in accordancewith one embodiment.

FIG. 20H is an isometric cutaway view of a processing system containinga specimen holder with a tissue sample in accordance with oneembodiment.

FIG. 20I is a side cross-sectional view of components of the processingsystem of FIG. 20H.

FIG. 20J is a schematic illustration of a monitoring system foranalyzing a tissue sample in accordance with one embodiment.

FIG. 20K is a front left side view of an analyzer in accordance with oneembodiment.

FIG. 20L is a front view of the analyzer of FIG. 20K.

FIG. 21A is an elevational view of a processing system capable ofperforming multiple treatments on tissue samples in accordance with oneembodiment.

FIG. 21B is a plot of frequency versus phase comparison in accordancewith one embodiment.

FIG. 21C is a plot of temperature versus time of flight in a media.

FIG. 21D is a plot of time versus time of flight for a tissue sample andtime of flight for media.

FIG. 21E is a plot of time versus a reference time of flight.

FIG. 21F is a plot of time versus a target time of flight.

FIG. 21G is a plot of time versus time of flight for a tissue sample.

FIG. 22 is an example process flow diagram illustrating an exampleworkflow variation between paper forms and e-LRF pen, illustratingexample information flow from pen to laboratory information system(LIS).

FIG. 23A is an exemplary sample vial.

FIG. 23B is an example vial along with an example specimen lid with clipand a cassette clipped to the lid.

FIG. 23C is another view of the cassette being inserted into the clipcoupled to the specimen lid.

FIG. 23D illustrates exemplary cassettes in open and closed positions.

FIG. 24 is an expected interactions process before processing.

FIG. 25 is an exemplary cold tissue kit including an exemplary box kitassembly, shown with, the specimen vials, specimen lids, a TEMPODcasing, and datalogger, separate, without being inserted yet into thefoam inserts, along with cassettes, according to an exemplaryembodiment.

FIG. 26A and FIG. 26B are an example operating room system workflow andan example AP Lab workflow, respectively.

FIGS. 27A and 27B illustrate exemplary sealing options and variationsfor an exemplary specimen cup, including a wrap cup in parafilm, and/orfume reduction concept including inserting a cassette through anexemplary septum of a membrane seal, respectively.

FIG. 28 is an illustration of an integrated cup design inship-to-hospital position.

FIG. 29 is an illustration of an integrated cup design illustrating areversed cap direction immersing a sample cassette in fixative in thevial.

FIG. 30A illustrates a method of use of the unit of FIGS. 29 and 30.

FIG. 30B illustrates a cassette with a slanted portion for labeling.

FIG. 30C is an illustration of samples loaded and ready for shipment toa lab.

FIG. 30D is a partial cutaway view of the image of FIG. 31C.

FIG. 30E is a protected version of the vial container.

FIGS. 31A-31C are individual components of a cassette holder, cassettereceiver, and fixative vial.

FIGS. 32A and 32B illustrate a user receiving the cassette holder, andthe cassette receiver/vial assembly, respectively.

FIGS. 33A, 33B, 33C, and 33D illustrate a process of attaching thecassette to the cassette holder; piercing a septum of the cassettereceiver/vial assembly; completely sealing a vial, ready for transport;and having the receiving lab unscrew the cassette holder/receiverassembly and removing the cassette, respectively.

FIGS. 34A and 34B illustrate a first state with a sample in the airexposed to the environment with a wedge arm that breaks a membrane; anda fixative vial with a breakable membrane, respectively.

FIGS. 34C and 34D illustrate a continuation of the first state of FIGS.34A and 34B with the sample container lid of FIG. 34A affixed to theprotective cylinder, with the fixative container of FIG. 34B stillisolated.

FIG. 34E illustrates a transition to a second state, by screwing thesample container around the fixative container, isolating the samplefrom the environment, with the fixative still isolated prior to the armbreaking the membrane.

FIGS. 35A-35I is an exemplary fume reduction feature explained, with anexemplary cassette cover in an exemplary dry implementation.

FIGS. 36A-36I is an exemplary preanalytical workflow.

FIGS. 37-44E illustrate example specimen vials, lids, clips, andcassettes, of various example embodiments.

FIG. 37 is an exemplary exploded view of an exemplary tissue pot withlid, and exemplary 10% NBF.

FIG. 38 is an exemplary exploded view of an exemplary tissue pot withlid, and exemplary 30% NBF.

FIG. 39A is a side view and FIG. 39B is a top view of an exemplaryCELLSTOR lid modification, of an exemplary specimen lid.

FIGS. 40A-40D are various views of an exemplary cell block cassetteclip, having an exemplary angled slot for clipping to an exemplarycassette.

FIGS. 41A-41D, and 42A-42 B are various views of an exemplary cell blockcassette.

FIGS. 43A-43C are various views of an exemplary cell transport capplate, configured as a cap for the specimen lid.

FIGS. 44A-44E are various views of an exemplary cell transport sealblock, which resembles a straight screw head, including an exemplaryslotted cassette clip of one embodiment, as can be used in the processset forth in FIGS. 35A-35I.

FIGS. 45A-45C are digital images of a transport device that isespecially configured for an autonomous warming embodiment.

FIGS. 46A-46C demonstrate the results of an experiment with anautonomous heating device as depicted herein stored at a variety ofambient temperatures. FIG. 46A is a graph of the temperature of thesample container under various ambient temperature conditions. FIG. 46Bis a series of digital images of tissues immunohistochemically stainedfor pAkt after the experiments. FIG. 46C is a bar graph of the relativestain intensity from tissues immunohistochemically stained for pAkt.

FIGS. 47A-47D illustrate exemplary embodiments of a simple timer usefulwith the present invention.

DETAILED DESCRIPTION OF THE INVENTION

Formalin fixation is a mainstay of histopathologic analysis, yet thepractice is poorly standardized and a significant potential source ofpre-analytical errors. Concerns of workflow and turnaround time drivedevelopment of shorter protocols, but abbreviated protocols lead to poorhistomorphology or inadequate downstream assay results. Additionally,immunohistochemistry assays for phosphorylated epitopes or otherbiomarkers have been challenging in the context of formalin fixedtissue. Standardized fixation parameters issued for clinical biomarkerassays to mitigate the errors from pre-analytical variation, such as theASCO/CAP guidelines for HER2 IHC, call for fixation in neutral bufferedformalin for 6 hours to 72 hours. We studied basic formalin biochemistryto develop a formalin fixation protocol that involves a pre-incubationin sub-ambient temperature formalin prior to a brief exposure to heatedformalin. This protocol is more rapid than standard protocols yetpreserves histomorphology and biomarkers such as phosphorylatedproteins, mRNA, and miRNA, for example. We have determined thatbiomarker preservation for extended times is compatible with possibletissue transport solutions.

Commonly owned patent application US 2012/0214195 A1 discloses atwo-temperature, rapid formalin fixation protocol that reduces fixationtime and is optimized for use across a broad range of tissue typesthrough better preservation of histomolecular features. Thetwo-temperature process involves soaking the tissue specimen inprecooled formalin for up to five minutes and then letting the samplesoak for two hours at 4° C., and another two hours at 45° C., accordingto an exemplary embodiment. This can be generally performed on standardlaboratory equipment such as an ice bucket or heating plate, in anexemplary embodiment. Finally, dehydration, clearing, paraffinization,embedding and sectioning are generally performed in an automaticstandalone tissue processor (such as, e.g., but not limited to, Leica,Sakura, Avantik, etc.), according to an exemplary embodiment.

The inventors have devised an integrated specimen collection andtransport solution that ensures specimens are optimized for subsequentstaining and analysis using a cold packing method that can include thefirst half of the two-temp fixation process, according to an exemplaryembodiment. By the time the pack is opened at the anatomical pathology(destination) laboratory, the specimen has been properly fixed at theappropriate cold temperature, according to an exemplary embodiment. Thespecimen can then be heated up, using, e.g., but not limited to,standard laboratory equipment or specialized Ventana hardware, etc., forthe requisite two hours, and can be subsequently ready for tissueprocessing, according to an exemplary embodiment.

The integrated system can also provide digital tracking, in the form ofspecimen temperature and time recording that can enable pathologyprofessionals to monitor conditions that may affect subsequentprocessing or analysis, according to an exemplary embodiment. The systemis designed for use with either paper-based laboratory request forms, ora digital e-LRF pen that accompanies the specimen throughout its journeyand allows for paperless routing, as well as enhanced specimen trackingthat can be synced to wireless devices or uploaded for cloud computing,according to an exemplary embodiment.

Some of the problems with conventional best-practice histology sample,collection, and transport methodologies include the following:

Current room-temperature fixation protocols do not preserve biomarkerssufficiently.

Tissue storage after removal is not sufficiently standardized withrespect to biomarker preservation; this can result in some specimenseven being left without formalin over the weekend in a refrigerator. Thelack of rigid adherence to set protocols across institutions and sitesoften results in widely varying degrees of histomolecular degradation.This compromises downstream processing, especially in samples subjectedto a variety of biomarker-based diagnostics.

Fixative fluid spills as a result of inversion or jostling.

Variations in ambient temperature during transport, which has been knownto affect efficacy of fixative.

Operator exposure to caustic materials and fumes during handling.

Exemplary problems with conventional standardized tracking methodologiesinclude:

General best-practices may vary intra- and inter-institutionally inspite of barcode labeling.

Current methodologies rely heavily on paper and individualized computerinput which may also vary from institution to institution; chain ofcustody is established through a series of signatures.

Real-time tracking of the computer specimen is paper-based and cannotprovide an up-to-the-minute accounting of temperature, location andtransport time.

The end result of these challenges is questionable sample quality andbiomarker preservation may not correctly reflect the original state ofthe tissue and lack of traceability during processing. These areunacceptable risks when dealing with oncology patients.

We have devised an integrated specimen collection and transport solutionusing a cold storage/transport method comprising: (a) a sample containerassembly for holding sealing the tissue sample in an enclosedenvironment while immersed in a cold formalin-based fixative solution(preferably at a temperature in the range of 0° C. to 7° C.); (b) atemperature-responsive element capable of tracking and recording thetemperature of formalin-based fixative solution disposed in the samplecontainer over a period of time; and (c) a case comprising a cavitydefined by a material comprising an insulator and/or a cooling element.The sample container is inserted into the cavity of the case. Sufficientinsulator and/or cooling element is provided such that the temperatureof the formalin-based fixative solution is held at a temperature in therange of 0° C. to 10° C. (preferably from 0° C. to 7° C.) for at least 2hours at ambient temperature (i.e. from 15 to 40° C.). The temperatureresponsive element is positioned such that it records one or more of thefollowing data points

-   -   the temperature of the formalin-based fixative solution over a        period of time;    -   the amount of time that the formalin-based fixative solution is        below and/or above a predetermined temperature;

Optionally, a time may be provided that records the amount of time thatthe tissue sample has been exposed to the formalin-based fixativesolution. Other monitors may be provided to record data including:exposure of the device or system to an impact; opening of the deviceduring shipment and/or storage; leakage or loss of the formalin-basedfixative solution; and/or time of flight of acoustic waves through thetissue sample as a measure of diffusion of fixative.

As used herein, the term “temperature-responsive element” or“temperature-responsive assembly” shall include any device, apparatus,chemical, or other element that can track and record a temperature of amedium. Examples include thermometers, infrared temperature sensors,phase change materials (such as the WARMMARK time-temperature tag).

In some embodiments, the system uses prefilled specimen containersstored at reduced temperatures, a transport box with or without coldelements and a data logger for sample quality checking. Tissue samplescan also be pre-soaked by submerging them in pre-cooled fixative. Thepre-soaking process can occur while the tissue samples are transportedto reduce overall processing times and to enable fast and accuratemedical diagnosis. The pre-soaking process can be monitored with varioustypes of sensors, monitors and data loggers to obtain information aboutthe pre-soaking process or other information used for subsequent tissueprocessing, such as a histological processing (e.g., fixing, embedding,dehydrating, infiltrating, embedding, sectioning, and/or staining) Theinformation obtained from the sensors/monitors in the data logger aboutthe pre-soaking process can be used to determine, for example, (1)diffusion status of tissue samples, (2) target temperatures forfixation, (3) tissue processing times (e.g., when to remove the tissuesamples from fixatives). After fixing the tissue sample, the tissuesample can be removed from the fixative and can be subjected to one ormore histological processes performed based, at least in part, oninformation obtained from the monitoring.

To minimize or limit fixation during shipping of the tissue sample(e.g., shipping from a collection site to a processing site), thepre-cooled fixative can be at a pre-soaking temperature for a pre-soakperiod of time. The pre-soak process can involve diffusion of coldfixative throughout substantially the entire thickness or cross sectionof the tissue sample. After desired diffusion is achieved, the tissuesample can be heated to a fixation temperature that is higher than thepre-soak temperature to start and/or promote cross-linking. Byinhibiting or preventing fixation during transport, the fixation processcan be performed at the processing site to accurately control thefixation process. If the fixative is formaldehyde, cross-linking canoccur between formaldehyde molecules and amine containing cellularmolecules without significantly compromising the tissue characteristics(e.g., antigenicity, morphology, or both).

For processing a tissue sample in accordance with one embodiment.Generally, tissue samples are monitored from sample collection throughprocessing. Collected information can be used to perform subsequenthistology processing and to generate reports (e.g., reports used fordiagnosis, patient monitoring, billing, etc.), an audit trail (e.g., anaudit trail of specimen handling steps), a processing parameter log(e.g., a log that could be printed and as a quality record at the end ofthe processing), or the like.

Initially a tissue sample is taken from a subject and can be one or moresections of tissue, an organ, a tumor section, a smear, a frozensection, cytology prep, or cell lines. An incisional biopsy, a corebiopsy, an excisional biopsy, a needle aspiration biopsy, a core needlebiopsy, a stereotactic biopsy, an open biopsy, or a surgical biopsy canbe used to obtain the sample. In some embodiments, the tissue sample cancontain genomic DNA, RNA (including mRNA & miRNA), protein,phosphorylated protein or combinations thereof. Example tissue samplesinclude, but are not limited to tissue biopsy, pap cytological material,surgical specimen, amniocentesis samples and autopsy material. In oneexample, a sample includes a biopsy of an adenocarcinoma, a sample ofnoncancerous tissue, and a sample of normal tissue (from a subject notafflicted with a known disease or disorder).

The tissue sample is then placed in a transporter system with a media. Afreshly removed tissue sample can be pre-soaked by placing the tissuesample in fixative within an appropriate amount of time to prevent orlimit an appreciable amount of degradation (e.g., ischemia). In someembodiments, the tissue sample is taken from a subject and placed in thefixative within a relatively short amount of time, for example, lessthan about 1 minute, 2 minutes, 5 minutes, 30 minutes, 1 hour, 2 hours,or the like. In some embodiments, the fixative can bepre-chilled/pre-cooled to a temperature below room temperature down toabout 0° C. The tissue sample can be placed into an open specimencontainer containing the pre-cooled liquid fixative. Alternatively thetissue sample can be placed in a tissue cassette such as those known inthe art. The tissue cassette can then be placed in the specimencontainer to submerse the tissue in the pre-cooled liquid fixative. Thecomposition and temperature of the fixative can be selected to achieve adesired rate of diffusion and/or cross-linking. The fixative can beformalin solutions of formaldehyde in water used for preservation ofbiological specimens. If formalin used as a fixative it is typically 10%NBF, but other solution concentrations (e.g., 10%-80% NBF) also can beused. The tissue sample can remain immersed in the fixative while it isconveniently transported to another location.

Exemplary transporter systems can include, but are not limited to,packaging, a bottle, a vial, or other object used to hold liquid mediaand at least one tissue sample. The transporter systems can includemachine-readable code (e.g., optical symbology, magnetic pattern orelectromagnetic, or electrostatic signal having information content)that may relate to sample identity, patient information, sample origin,sample chain of custody, instructions for processing samples,information regarding the characteristics of samples, test results forsamples, images of samples, or other information associated with thetissue sample.

The fixative can be at a pre-soak temperature (or “diffusiontemperature”) to allow diffusion of the fixative while inhibitingfixation. The pre-soak temperature can be about 0° C. to about 15° C.,about 4° C. to an upper temperature about 10° C. or from about 3° C. toabout 5° C. For some procedures, the pre-soak temperature can be about4° C. Although some fixing (e.g., cross-linking) may occur during thepre-soaking process, the fixing primarily occurs after the pre-soakingprocess (i.e., during the fixation process). The pre-soaking process canbalance the beneficial properties associated with substantially completediffusion while minimizing or limiting the effects associated withinitializing or promoting cross-linking and preventing ischemia. In someembodiments, the rate of diffusion of the fixative is be maximized whilelimiting and minimizing any deleterious effects associated withincreased cross-linking rate. In other embodiments, the transportersystem can include room temperature fixative that is quickly chilled toinhibit ischemia and cross-linking while promoting fixative diffusioninto the sample.

The transporter system can be transported to an onsite laboratory or anoffsite site laboratory while the fixative diffuses through the tissuesample. The fixative can be kept at or below the pre-soak temperature,thereby allowing for a wide range of delivery times withoutsignificantly impacting the state of the tissue specimen. In someprocedures, the fixative is formalin with a target transport period oftime in a range of about 15 minutes to 72 hours or longer, e.g. 14 days.If the sample arrives at the processing site before completion of thetarget pre-soak period of time, the tissue sample can be left in thefixative to complete the diffusion process. Alternatively, the tissuespecimens can be removed from the transporter system and the pre-soakingprocess can be completed with another fixative.

Once at the laboratory the tissue sample can either be evaluated todetermine whether the fixative has adequately infused the sample tissueof the data compiled from the monitoring system can by analyzed todetermine the diffusion. After desired diffusion is achieved, thefixation process can be performed by, for example, heating the tissuesample to a fixation temperature to start and/or promote cross-linkingif the rate of cross-linking is temperature dependent. In someprocedures, the tissue sample can be removed from the transporter systemand delivered to a processing apparatus/system that can perform afixation process by immersing the tissue sample in warm fixative. If thefixative is formaldehyde, the temperature of the warm formalin can begreater than the ambient temperature and up to at least 55° C., moretypically from about 35° C. to about 45° C., as this temperature rangemay increase the cross-linking kinetics sufficiently to allow relativelyquick tissue cross-linking. However, if the temperature is increasedabove about 50° C., the tissue sample may begin to degrade, which mayhave a deleterious effect on certain subsequent histological reactions.Thus, the upper temperature and time period of the fixation process canbe selected to allow subsequent imaging processes, such as in situhybridization, IHC, and/or hematoxylin and eosin (H&E) processing. Thetime period for the fixation process can range from greater than about15 minutes up to at least about 5 hours, more typically is at leastabout 1 hour to about 4 hours, and more typically is from about 2 hoursto about 3 hours. In certain embodiments, the fixation process can beperformed for about 1.5 hour at a temperature of about 45° C. Thepre-soak fixative and the cross-linking fixative can be the same ordifferent. As yet another example, entirely different aldehydefixatives, such as formaldehyde and glutaraldehyde, can be used for thepre-soak process and fixation processes. Additionally, instead ofremoving the sample from the fixative, the pre-soak fixative can beheated to cause cross-linking. As such, the tissue sample can remainimmersed in the same fixative throughout the pre-soak and fixationprocesses.

Various factors may be considered to determine pre-soaking and fixationprocessing. These factors can include: sample thickness, which typicallyranges from about 1 mm to about 10 mm thick, more typically from about 2mm to about 8 mm thick, and even more typically from about 4 mm to about6 mm thick; volume of fixative to tissue sample mass, which typically isfrom about 10:1 to about 50:1 volume to mass; fixative composition;temperature; and sample immersion time in the fixative. Processingtimes, fixation history, condition history, tissue characteristics, orother histology information can be used to adjust processing to ensureproper tissue processing.

FIG. 1 is a workflow 190 for tracking tissue samples from a collectionsite to a processing site. At 200, a tissue sample can be taken from asubject at an operating room and loaded into a specimen container. Thespecimen container can be assigned identification information and loadedinto shipping container, such as a transporter system. At 204, thetissue sample is shipped from the collection site to the processing site(such as an onsite or offsite pathology lab) by, for example, a courieror a runner. The transport time can be less than 24 hours or as much as14 days. At 206, information (e.g., temperature condition, temperatureversus time information, diffusion state, etc.) from a monitoring systemof the transporter system can be used to determine an appropriatefixation protocol. The tissue sample can be fixed, and the fixed tissuesample can be processed using standard tissue processing techniques.

FIG. 2 is an exploded isometric view of a transporter system 400. Thetransporter system 400 includes a transporter container 404 having amain body 410 and a closure 412. The closure 412 can be separated fromthe main body 410 to access the specimen containers 422 (individually422 a, 422 b, 422 c, 422 d).

Referring to FIG. 2, specimen containers 422 can be similar one anotherand, thus, the description of one specimen container applies equally tothe others. The specimen containers 422 can be bottles, vials, pots, orother types of containers made of plastic, glass, or other materialsuitable for contacting fixatives. One example of such a container canbe CELLSTOR POT available from CellPath Ltd. (Newtown, Powys, UK)another BIOPSAFE is available from Axlab Innovation A/S (Vedbaek,Denmark). Other types of specimen containers can also be used. Thespecimen containers 422 a, 422 b, 422 c, 422 d can be held inreceptacles 432 a, 432 b, 432 c, 432 d of a tray 436. The tray 436 canbe a solid metal tray with a relatively high thermal mass capable ofabsorbing significant amounts of heat. The tray 436 can be cooled in afreezer unit and placed into the main body 410, which can be at roomtemperature. The containers 422 can be inserted the tray 436, whichabsorbs heat from the containers 422 and their contents. Alternatively,the rack 436 can be made, in whole or in part, of one or more thermallyisolating materials, such as open-cell foam or closed cell foam, tofurther increase storage time of the pre-cooled specimen containers 422.Additionally, thermally isolating materials may provide anti-freezeprotection for the sample in adverse transport conditions (e.g., duringcold winter conditions). The temperature strip 423 can be used tomonitor the temperature of the tray 436.

A monitoring system 450 can obtain and store information about thetissue samples in the respective specimen containers 422 and includesdetection devices 442 (individually 442 a, 442 b, 442 c, 442 d) and acontroller 450. In some embodiments, the detection devices 442 includeradio-frequency identification (RFID) tags with a small metallic antennaand a silicon chip, and can be active or passive. The informationcontent of an RFID tag can be fixed or changeable. The RFID tags cancommunication information to the communication device 452. Additionallyor alternatively, the detection devices 442 can include sensing elements(e.g., temperature sensor that measure the measure a temperature of thetissue sample) or can be in communication with sensing devices carriedby the containers 422. The communication device 452 can be in the formof an RFID code reader, which are well known in the art and typicallyinclude an antenna and a transceiver that receives information from theRFID tag. Because the communication device 452 is physically coupled tothe container 410, the communication device 452 can remain incommunication with the tag throughout shipping.

Monitoring system 450 can also comprise real time monitoring of themovement of the fixative through the tissue sample. The composition ofthe fixative may be selected to enhance monitoring. For example, NBF hasa relatively high bulk modulus compared to interstitial fluid. The soundtransmissibility of the fixative is related to its bulk modulus ( ) anddensity ( ) according to the speed equation, speed of sound infixative=. The fixative, such as formalin, with a bulk modulus greaterthan interstitial fluid can significantly alter the TOF as it displacesthe interstitial fluid.

A TOF acquisition scheme may be used to monitor tissue samples. The TOFacquisition scheme may include an A/D conversion scheme (e.g., about 1μsec phase comparison) to obtain a large number of phase comparisons toprovide generally real-time monitoring. The phase comparisons may beperformed at the same frequency and phase relationship, and thetemperature of the fixative and/or tissue sample can remain generallyconstant to increase signal to noise ratios. Because fluctuations intemperature may cause measureable changes in TOF, the TOF acquisitionscheme can compensate for changes in TOF attributable to, for example,temperature changes.

In addition to acoustic monitoring the monitoring system canadditionally be comprised of one or more sensors selected to, amongother capabilities, determine, monitor or store data about: thetemperature of the container and/or sample; elapsed time; environmentaltemperature in the transporter; ambient light to determine if thecarrier assembly and sample container are in or outside the transporter;infrared proximity (determine if specimen or a cassette containing thespecimen is in the specimen container or the container is in the carrierassembly, etc.); accelerometer to records impact or inversion; leaks orfluid loss; or any other type of sensors or sensor arrays. Additionally,one or more sensors may be configured to transmit data, such astemperature, for example, while being submerged in the reagent in thespecimen holder in the reagents.

FIG. 3 is a cross-sectional view of a transporter system 2200.

FIG. 3 depicts an illustration 2200 of an exemplary four cup transportsystem. The exemplary cold tissue transport system, as illustrated inFIG. 3 can maintain a temperature of the specimen within an exemplaryrange of 2-8° C. by using two exemplary elements: vacuum insulationpanels 2204, 2222, and phase change blocks 2206, 2208, 2214, 2216. Thebox is assembled as shown in FIG. 3. According to exemplary embodiment,an exemplary strap, such as, e.g., but not limited to, a coupler, suchas, a VELCRO strap, can hold an exemplary vacuum insulation panel lid2204, onto a vacuum insulation box 2222 in exemplary outer box 2220, anexemplary chill −20° C. block 2206, another exemplary chill 4° C. block2208, an exemplary foam insert 2210, to which the exemplary strap 2202can be coupled, an exemplary foam insert 2212 can be configured toreceive exemplary specimen containers 2218, as shown, and exemplarychill 4° C. block 2214, and the other exemplary chill −20° C. block 2216can be placed beneath the foam insert 2212, according to an exemplaryembodiment.

The exemplary vacuum insulation panels 2204, 2222 can provide excellentinsulation from the environment, according to an exemplary embodiment.The exemplary panels 2204, 2222 line all sides of the interior of thebox 2220 with a fairly tight fit, according to an exemplary embodiment.The exemplary insulation panels 2204, 2222, along with the phase changechill or ice blocks 2206, 2208, 2214, 2216, nest the specimens tightlyin their exemplary carrier assemblies 2218, which can be seated, asillustrated in 2200, into an exemplary foam rack 2212, 2210, holding anexemplary four cups 2218.

The exemplary phase change blocks 2206, 2208, 2214, 2216 can providethermal buffering from the environment, according to an exemplaryembodiment. Two types of blocks can be used simultaneously to maintainthe sample temperature between 2° C. and 8° C., according to anexemplary embodiment. The exemplary green blocks 2206, 2216 can beprepared in a −20° C. freezer (at least overnight) and can be placedtoward the outside of the shipping box, away from the foam inserts 2210,2212 which hold the samples, as illustrated in FIG. 3. In order toprovide proper thermal buffering, the green blocks' 2206, 2212temperature should be no lower than −20° C. (if it is lower it mayfreeze the sample) and no higher than −15° C. (if it is higher it won'tlast as long), according to an exemplary embodiment. The green blocks2206, 2212 should not be placed in a freezer with a defrosting function,according to an exemplary embodiment. The exemplary orange blocks 2208,2214 can be prepared in a 4° C. refrigerator (at least overnight) andcan be placed next to the foam inserts 2210, 2212 which can hold thesamples, maintaining them at 4° C., according to an exemplaryembodiment. Prior to packing the box 2220, the orange blocks 2208, 2214should be removed from the refrigerator; if the blocks 2208, 2214 arecompletely frozen, the blocks 2208, 2214 should be allowed to warm untilmost or all of the blocks' 2208, 2214 content is liquid, according to anexemplary embodiment. If the blocks 2208, 2214 are either partially orcompletely liquid and still at 4° C., the blocks 2208, 2214 are readyfor use, according to an exemplary embodiment.

Carrier Assemblies (CryoPack carrier assembly FIG. 7A-G, and TEMPODcarrier assembly FIG. 8A-H)

FIG. 4A illustrates an exemplary carrier assembly 2218 including anexemplary carrier assembly 2404 shown including an exemplary TEMPODcarrier assembly 2404 with an exemplary specimen vial 2302 with specimenlid 2402 atop it, and an exemplary data logger 2406, inserted inexemplary slots therein, according to an exemplary embodiment. ExemplaryTEMPOD carrier assembly 2404 can be an exemplary plastic casing that canseat the vial 2302 with lid 2402, and data logger 2406, which togethercan be placed into the Styrofoam inserts 2210, 2210 inside the GreenBox2220, according to an exemplary embodiment.

As previously mentioned the monitoring assembly may include one or moresensors selected to, among other capabilities, determine, monitor orstore data about: the temperature of the container and/or sample;elapsed time; environmental temperature in the transporter; ambientlight to determine if the carrier assembly and sample container are inor outside the transporter; acoustic; infrared proximity (determine ifspecimen or a cassette containing the specimen is in the specimencontainer or the container is in the carrier assembly, etc.);accelerometer to records impact or inversion; leaks or fluid loss; orany other type of sensors or sensor arrays. Additionally, one or moresensors may be configured to transmit data, such as temperature, forexample, while being submerged in the reagent in the specimen holder inthe reagents.

Data loggers (FIGS. 4B, 4C, 9A and B, and 10A and B)

FIG. 4B illustrates an exemplary embodiment of an exemplary currentimplementation including an exemplary TEMPOD data logger 2406, which canbe used to monitor the temperature of specimen samples in a vial 2302and can be used to maintain a chain of custody over the samples in anaccompanying associated vial 2302 with lid 2402, as shown in FIG. 4A. Inan example embodiment, the TEMPOD data logger 2406 can be used for bothcold- and room-temperature transport. The data logger 2406 of thecarrier assemblies 2218 record and monitor temperature excursionsagainst exemplary preprogrammed thresholds (e.g., but not limited to,heating, and/or freezing, etc.), about which can be indicated with anexemplary display such as, e.g., but not limited to, a LED/LCD displaywith exemplary warning(s). These exemplary data logger(s) 2406 can bepreconfigured for the first sample collection. Alternatively, asillustrated in FIG. 4C, Cryopak data loggers 2408 can also be used forboth cold- and room-temperature transport, according to anotherexemplary embodiment. Each of the data loggers 2406, 2408 can come withassociated accompanying, proprietary software, according to an exemplaryembodiment.

FIGS. 6A and 6B depict an exemplary exploded view 2700 and inserted view2710, respectively of a an exemplary specimen vial 2302, specimen lids2402, cassette 2304, 2306, coupled to lid 2402 by an exemplary clip (notlabeled, with various embodiments discussed herein) and carrier assembly2404, and CRYOPACK data logger 2408 (discussed herein with reference toFIG. 4C), according to an exemplary embodiment.

An exemplary embodiment can include data logging electronics 2406, 2408,which can be held separately from the vial 2302 via carrier assembly2404 construct, with thermal communication to allow monitoring by thedata logger 2406, 2408 of the vial and its sample contents.

An exemplary embodiment can provide an exemplary timer/clock functionassociated with the insertion of the container, and can log other timesand/or temperatures of events, or at specific and/or periodic times.

An exemplary embodiment can provide an exemplary feature having a meansto indicate completion of a predetermined time after the insertion ormanual start was established.

An exemplary embodiment can provide an exemplary display, monitor orindicator (such as, e.g., with an indicating/monitoring/displayingand/or storing system and/or method LED/LCD).

An exemplary embodiment can provide an exemplary feature having atemperature recording capability.

An exemplary embodiment can provide an exemplary feature of having ameans to record and indicate temperature excursions againstpreprogrammed thresholds (heating, freezing), indicating with LED/LCD.

An exemplary embodiment can provide an exemplary feature having a meansto monitor temperature excursions and time after sealing of thecontainer, and can indicate successful completion per LED/LCD againstpreprogrammed thresholds.

An exemplary embodiment can provide an exemplary feature of generatingone or more of the warnings set forth in Table 1. Moreover, thetriggering of such a warning may instigate one or more follow-up actionsduring a shipping/fixation process as set forth in Table 1.

TABLE 1 Warning Risk/Follow-up Action Temperature exceeds Potential lossof labile markers; verify threshold that occurrence was related tocritical phase of surveillance profile Temperature falls Potential lossof sample due to freezing below threshold with impact to morphology.Verify that fluid temperatures stayed above freezing point. Packageexperienced an impact Potential damage to hardware and/or exceeding athreshold sample integrity; test hardware to ensure proper function andexamine sample to ensure it remained sufficiently intact A leak or lossof Potential operator exposure to fixative fixative is detected andpotential impact to sample integrity (e.g. drying and/or unevenfixation) and; exercise caution upon handling and verify fluid contentsof container Undertime alarm Insufficient fixation; Continue to keepsample and datalogger in cold environment until minimum time has beenmet Overtime alarm Overfixation and/or loss of labile biomarkers; Reviewspecimen transport data and verify that no other conditions may havecompromised sample integrity.

An exemplary embodiment can provide an exemplary feature including oneor more memories to store and/or include details about the sample ID,location relative to Wi-Fi transponders, and/or record date/time, etc.

An exemplary embodiment can provide an exemplary means to communicatevia RFID/Bluetooth/NFC/Wireless/Zig Bee or direct connection to aPC/tablet/Cell Phone.

FIGS. 7A-G depict exemplary CRYOPACK carrier assemblies, according to anexemplary embodiment. FIG. 7A depicts a cutaway cross-sectionorthographic view 4500 of carrier assembly 2404. FIG. 7B depicts a leftside orthographic view 4510 of carrier assembly 2404. FIG. 7C depicts abottom side orthographic view 4520 of carrier assembly 2404. FIG. 7Ddepicts a front side orthographic view 4530 of carrier assembly 2404.FIG. 7E depicts a top orthographic view 4540 of carrier assembly 2404.FIG. 7F depicts a right side orthographic view 4550 of carrier assembly2404. FIG. 7G depicts an isometric perspective view 4560 of carrierassembly 2404.

FIGS. 8A-H are exemplary TEMPOD carrier assemblies, according to anexemplary embodiment. FIG. 8A depicts a bottom orthographic view 4600 ofcarrier assembly 2404. FIG. 8B depicts a front side orthographic view4610 of carrier assembly 2404. FIG. 8C depicts a top orthographic view4620 of carrier assembly 2404. FIG. 8D depicts a right side orthographicview 4630 of carrier assembly 2404. FIG. 8E depicts a back orthographicview 4540 of carrier assembly 2404. FIG. 8F depicts a cutawaycross-section orthographic view 4650 of carrier assembly 2404. FIG. 8Gdepicts a right side orthographic view 4660 of carrier assembly 2404.FIG. 7 depicts an isometric perspective view 4570 of carrier assembly2404.

FIG. 9A depicts an exemplary front view 4700 of an exemplary CRYOPACKdata logger 2408.

FIG. 9B depicts an exemplary side view 4710 of the exemplary data logger2408.

FIG. 10A depicts an exemplary front view 4800 of an exemplary TEMPODdata logger 2406.

FIG. 10B depicts an exemplary side view 4810 of the exemplary datalogger 2406.

Paper-Based LRF

FIG. 5A illustrates an example initial paper-based form, which can beincluded in the packet specifically for use with the SCTT system. TheSCTT forms are likely to be provided by an external vendor (PharmaFormsoffers an integrated solution that can be used for this kit) and/or acustom-designed form may be used with the Ventana SCTT solution,according to an exemplary embodiment. Software may be purchased, whichmay allow the forms to be both designed and printed by the customer,according to an exemplary embodiment.

Data collected may include, according to an exemplary embodiment:

At a beginning of surgery: patient ID, collection Site, responsibleparty, date of surgery.

1) As samples are collected: a sample identifier, a time of collection(beginning of ischemia), an anatomical site of collection, a type offixative and/or a fixative concentration.

2) Any comments or anomalies in the collection process may be noted. Thedate/time of sample shipment can be recorded.

Paper lab request forms (LRF) can be custom designed (using, e.g., butnot limited to, www.pharmaforms.com/en, etc.) for the cold tissuetransport method, or conventional LRFs may be used either alone, or,optimally, in concert with the eLRF digital pen as shown in FIG. 5B or5C, which can have Wi-Fi 33 and/or cloud computing capabilities (theexemplary form shown in FIG. 5 is a proprietary Roche LRF.

Digital pen-based eLRF (FIGS. 5B and C, and 11A and B)

FIGS. 5B and 5C depict various exemplary views of an exemplaryelectronic pen.

Electronic lab request forms (eLRF) can be custom designed, e.g., forthe cold tissue transport method, or conventional LRFs may be usedeither alone, or, optimally, in concert with the eLRF digital pen, whichcan have Wi-Fi 33 and/or cloud computing capabilities (the exemplary LRFform of FIG. 5A is an exemplary proprietary Roche LRF.

Example external vendors such as, e.g., but not limited to, PharmaFormcan also offer digital tracking pens (e.g., ANOTO Pen) based on opticalcharacter recognition technology, according to an exemplary embodiment.Similar to the form-design and -printing, process, several options existfor extracting the data from the example ANOTO Pen, according to anexemplary embodiment. The customer may use software on a local personalcomputer (PC) or other computing device to extract data from the digitalpen into a file which can then be emailed or uploaded to interestedparties, according to an exemplary embodiment. Alternatively, pen datacan be uploaded to a PharmaForms-provided cloud solution where the datacan be verified by a PharmaForms representative and then can be madeavailable for download by parties granted access to the site, accordingto an exemplary embodiment.

The pen may travel with the specimen sample containers and arrive at theoperating room at the same time they do, according to an exemplaryembodiment. When the samples leave the operating room and travel to thelab, the digital pen can go with the samples, according to an exemplaryembodiment. Alternatively, the pen may travel with the individual who isresponsible for collecting the samples, according to another exemplaryembodiment. When the samples leave the lab, the written form can travelwith the samples, but the pen can stay with its owner, according to anexemplary embodiment. The pen's electronic data can be uploaded by thepen's owner after the samples are enroute to the lab, according to anexemplary embodiment.

FIGS. 11A and 11B depict exemplary DIGITAL e-LRF PEN (various typespossible, including with PharmaForms), as may be used in variousembodiments of the claimed invention.

FIG. 11A is an example ANOTO branded digital pen with exemplaryfunctionality and components including, e.g., but not limited to, anexample wireless transceiver (e.g., BLUETOOTH, etc.) 4902, an examplebatter 4904, an example memory 4908, an example processor 4910, exampleink 4906, and an example camera 4912, according to an exemplaryembodiment.

FIG. 11B is an example diagram 4914 illustrating an example LOGITECH IObranded digital pen and components, with example components including:an example optical sensor 1, an example processor 2, an exampleuniversal serial bus (USB) cradle 3, an example ink cartridge 4, anexample memory 5 (e.g., for storing up to an example 40 pages), anexample battery 6 (e.g., lasting up to 25 pages of writing), and Capcontrols 7 (e.g., on/off), according to an exemplary embodiment.

FIG. 12 is an example process flow diagram 5000 illustrating an exampleworkflow variation between paper forms and e-LRF pen, illustrating formscreated and distributed by PharmaForms, according to an exemplaryembodiment.

Flow diagram 5000 can begin with 5002, where PharmaForms designs andprints forms. From 5002, flow diagram 5000 can continue with 5004.

In 5004, forms can be mailed to the responsible party. From 5004, flowdiagram 5000 can continue with 5006.

In 5006, the responsible party can arrive in the operating room withsample containers, paper forms, and their own digital pen. From 5006,flow diagram 5000 can continue with 5008.

In 5008, surgery is performed, and paper forms can be filled out using adigital pen. From 5008, flow diagram 5000 can continue with 5010, or5016.

In 5016, patient samples and paper forms can travel to the AP lab,according to one exemplary embodiment.

In 5010, the responsible partner can upload pen data, according to anexample embodiment. From 5010, flow diagram 5000 can continue with 5012.

In 5012, an AP lab can receive the sample data before samples arrive,according to an example embodiment. From 5012, flow diagram 5000 cancontinue with 5014.

In 5014, samples arrive at the AP lab. From 5014, flow diagram 5000 canend, according to an example embodiment.

FIG. 13 is an example process flow diagram 5100 illustrating an exampleworkflow variation between paper forms and e-LRF pen, illustrating formscreated by a responsible party, according to an exemplary embodiment.

Flow diagram 5100 can begin with 5102, where a responsible party designsand prints forms according to an example embodiment. From 5102, flowdiagram 5100 can continue with 5104.

In 5104, the responsible party can arrive in the operating room withsample container forms can be mailed to the responsible party, accordingto an example embodiment. From 5104, flow diagram 5100 can continue with5106.

In 5106, surgery can be performed, according to an example embodiment.Paper forms can be filled out using an example digital, according to anexample embodiment. From 5106, flow diagram 5100 can continue with 5108or 5114.

In 5108, the responsible party can upload the pen data, according to anexample embodiment. From 5108, flow diagram 5100 can continue with 5110.

In 5110, the AP lab can receive the sample data before samples arrive,according to an example embodiment. From 5110, flow diagram 5100 cancontinue with 5112.

In 5112, the samples can arrive at the AP lab, according to an exampleembodiment. From 5112, flow diagram 5100 end, according to an exampleembodiment.

In 5114, patient samples and paper forms can travel to AP lab, accordingto an example embodiment. From 5114, flow diagram 5100 can continue with5112, according to an example embodiment.

FIG. 14 is an exemplary diagram 5400 illustrating an exemplary samplecollection and tracking system (SCTS), according to one embodiment.Diagram 5400 can include a clinical sample operations (CSO) component5402 including LIMS 5440, coupled to IT/Data Management database (DB),which includes sample tracking concepts, i.e., traveling pen, tablet,etc., receives data from specimen collection management 5412, fromprefilled container vial 2302, with integrated cassette 2304, 2306(avoiding barcode or RFID), a cold storage box 5416, loading monitor,data logger 5426, transport 10-72 hours, accessioning managementsoftware/user interface 5422, transport/integrity, 5424, standard TP5430, an embedded block 5432, a clinical data capture 5406 with clinicaldata, and clinical databank 5428, and in the Laboratory 5405, a LIMS5434, a vantage process 5436, a slide review and interpret process 5438using cutting and staining, according to an exemplary embodiment.

FIG. 15 is an example diagram 5600 of tracking via an example electronicpen. As illustrated various processing can be provided by tracking viaelectronic pen. The exemplary, but non-limiting process illustrated caninclude: using an ePen to fill out the lab requisition form (LRF) andcan collect electronic data; collecting barcode information from apatient's wristband, container/vial, and cassette; optionallytransferring by ePen electronic data to a computing device (e.g.,tablet, mobile phone, etc.) for verification and/or correction; all ofwhich can be in the surgical suite/collection site, in an exampleembodiment. Optionally, the verified electronic data can be uploaded viaweb/cloud/mobile phone/etc. to LIS, e.g., the pen can stay onsite orwith the same user, in an example embodiment. Further, in an exampleembodiment, the ePen can travel with the specimen in the same coolingbox 2200, if upload is not possible. Further, in an example embodiment,the ePen can be received at the accessioning stage in the histology labfor exemplary readout and/or reporting to the LIS; and if neededcorrections may be made using, e.g., the paper form, etc. In an exampleembodiment, the data logger 2406, 2408 can be readout and integratedinto the data uploaded to the LIS.

The process can include, establishing chain-of-custody during specimencollection, integrating data such as patient-ID, location of surgery,day/time, name of surgeon, location of specimen removal; associatingwith a designated container/cassette barcode ID, recording the insertiontime of the specimen into the fixative vial, logging the transportconditions (temp/time profile); and documenting a total time of colddiffusion after which the sample can be safely removed and submitted toa heated formalin step for crosslinking: duplicate capture ofhandwritten Lab Requisition Form by means of Electronic Data Capture andassociating it to a specific Specimen or Cassette via Barcode/RFID,verifying minimum fixation conditions; and/or enabling such data capturefor a Clinical Trials, according to an example embodiment.

According to an exemplary embodiment, identifier use processing caninclude use of a unique identifier (Anoto code) printed onto the labelof the container and being part of the capture of the eLRF, including,e.g., the barcode on the cassette could be, i.e., a Vantage compatiblepreprinted code, that is being inserted into the container; and/or whenthe container arrives at the accessioning stage, the cassette barcodecan be identified by a barcode reader and tied to the patient-ID thatwas previously associated to the container (Anoto) code, according to anexample embodiment.

According to an exemplary embodiment, processing can include use of thecontainer (pot) also having a unique label with Anoto pattern (in littlesquare box), including, e.g., the container can still be pickedrandomly, and the pen can tie that code electronically to thepatient-ID., and can also need a handwritten link as well on paper (someID that they have to copy from the pot's label onto paper), according toone example embodiment.

FIG. 16 is an example diagram 5700 of tracking via an example electronictablet device. As illustrated various processing can be provided bytracking via tablet or other portable computing or communicationsdevice. The exemplary, but non-limiting process illustrated can include:using a tablet in capturing pertinent information about the patient andstudy (duplicate entry of some of the lab requisition form (LRF)) and/orto fill out the lab requisition form (LRF) and can collect electronicdata; collecting barcode information from a patient's wristband,container/vial, and cassette via, e.g., an external reader, anapplication (“App”), etc. etc.; optionally transferring by tabletelectronic data to a computing device (e.g., tablet, mobile phone, etc.)for verification and/or correction; data entry errors can be corrected,even on screen, if needed; where all of which can be in the surgicalsuite/collection site, in an example embodiment. Optionally, theverified electronic data can be uploaded via web/cloud/mobile phone/etc.to LIS, e.g., the pen can stay onsite or with the same user, in anexample embodiment. Further, in an example embodiment, the tablet cantravel with the specimen in the same cooling box 2200, if upload is notpossible. Further, in an example embodiment, the tablet can be receivedat the accessioning stage in the histology lab for exemplary readoutand/or reporting to the LIS. In an example embodiment, the data logger2406, 2408 can be readout and integrated into the data uploaded to theLIS.

The process can include, establishing chain-of-custody during specimencollection, integrating data such as patient-ID, location of surgery,day/time, name of surgeon, location of specimen removal; associatingwith a designated container/cassette barcode ID, recording the insertiontime of the specimen into the fixative vial, logging the transportconditions (temp/time profile); and documenting a total time of colddiffusion after which the sample can be safely removed and submitted toa heated formalin step for crosslinking: duplicate capture ofhandwritten Lab Requisition Form by means of Electronic Data Capture andassociating it to a specific Specimen or Cassette via Barcode/RFID,verifying minimum fixation conditions; additional data capture ofbarcodes via camera or RFID/NFC tags via built in communicationinterface; additional capture of image of surgical site; and/or enablingsuch data capture for a Clinical Trials, in one example embodiment.

Exemplary Use of Wi-Fi MAC Address for Tracking

According to an exemplary embodiment, a device capable of having a MACaddress and Wi-Fi communication could be used as a data logger andpositional time stamper for sample tracking. According to an exemplaryembodiment, any time either the Wi-Fi router or MAC addressed devicecommunicated, the identity of both could be logged. Since the time ofcontact is known and the location of Wi-Fi router is known, one coulduse the massive network of free-access Wi-Fi routers to record thelocation of the MAC device throughout its journey from point A to pointB, according to an exemplary embodiment.

FIG. 17 is an example diagram 5800 of tracking via an example staticcomputer at the collection site. As illustrated various processing canbe provided by a stationary, i.e., static computing, or portablecomputing or communications device. The exemplary, but non limitingprocess illustrated can include: using a computer using a computer,e.g., PC, in capturing pertinent information about the patient and study(duplicate entry of some of the lab requisition form (LRF)) and/or tofill out the lab requisition form (LRF) or equivalent data, and cancollect electronic data; collecting barcode information from a patient'swristband, container/vial, and cassette via, e.g., an external reader,etc.; optionally transferring by internet, electronic data, etc., to acomputing device (e.g., tablet, mobile phone, etc.) for verificationand/or correction; data entry errors can be corrected, even on screen,if needed; where all of which can be in the surgical suite/collectionsite, in an example embodiment. Optionally, the verified electronic datacan be uploaded via internet/web/cloud/mobile phone/etc. to LIS, e.g.,the computer can stay onsite or with the same user, in an exampleembodiment. Further, in an example embodiment, the computer can travelwith the specimen in the same cooling box 2200, if upload is notpossible. If LIS connectivity is prohibited, the computer could programthe data logger to retain additional information, etc. Further, in anexample embodiment, the computer can be received at the accessioningstage in the histology lab, where barcode information can be can be usedto reference the sample via LIS query, or for exemplary readout and/orreporting to the LIS. In an example embodiment, the data logger 2406,2408 can be readout and integrated into the data uploaded to the LIS, inone example embodiment.

The process can include, establishing chain-of-custody during specimencollection, integrating data such as patient-ID, location of surgery,day/time, name of surgeon, location of specimen removal; associatingwith a designated container/cassette barcode ID, recording the insertiontime of the specimen into the fixative vial, logging the transportconditions (temp/time profile); and documenting a total time of colddiffusion after which the sample can be safely removed and submitted toa heated formalin step for crosslinking: duplicate capture ofhandwritten Lab Requisition Form by means of Electronic Data Capture andassociating it to a specific Specimen or Cassette via Barcode/RFID,verifying minimum fixation conditions; additional data capture ofbarcodes via camera, or radio frequency identifier (RFID)/near fieldcommunication (NFC) tags via built in communication interface;additional capture of an image of surgical site using webcam, etc.;and/or enabling such data capture for a Clinical Trials, in one exampleembodiment.

FIG. 18 is an example diagram 5900 of tracking via an example travellingcooling/shipping box with intelligence. As illustrated variousprocessing can be provided by a travelling, cooling, shipping box withintelligence, i.e., movable/portable/mobile computing, or portablecomputing or communications device, etc. The exemplary, but non-limitingprocess illustrated can include: using a computer, e.g., PC, incapturing pertinent information about the patient and study (duplicateentry of some of the lab requisition form (LRF)) and/or to fill out theLRF, and can collect electronic data; collecting barcode informationfrom a patient's wristband, container/vial, and cassette via, e.g., anexternal reader, etc.; any data entry errors can be corrected, onscreen, if needed, optionally transferring by computer the electronicdata to a computing device (e.g., tablet, mobile phone, etc.) forverification and/or correction; data entry errors can be corrected, evenon screen, if needed; where all of which can be in the surgicalsuite/collection site, in an example embodiment. The PC can interfacewith a host computer in a storage box, etc. to store the data, in anexample embodiment. Loading/unloading of samples can be monitored by thehost computer built into the cooling box, in the example embodiment,together with logging of times and temperatures. At the accessioningstage, in the histology lab, the host computer can provide allelectronic data. Optionally, the verified electronic data can beuploaded via internet/web/cloud/mobile phone/etc. to LIS, e.g., thecomputer can stay onsite or with the same user, in an exampleembodiment. Further, in an example embodiment, optionally the computercan travel with the specimen in the same cooling box 2200, if upload isnot possible. Optionally, if LIS connectivity is prohibited, thecomputer could program the data logger to retain additional information,etc. Further, in an example embodiment, the computer can be received atthe accessioning stage in the histology lab, where barcode informationcan be can be used to reference the sample via LIS query, or forexemplary readout and/or reporting to the LIS. In an example embodiment,the data logger 2406, 2408 can be readout and integrated into the datauploaded to the LIS, in one example embodiment.

The process can include, establishing chain-of-custody during specimencollection, integrating data such as patient-ID, location of surgery,day/time, name of surgeon, location of specimen removal; associatingwith a designated container/cassette barcode ID, recording the insertiontime of the specimen into the fixative vial, logging the transportconditions (temp/time profile); and documenting a total time of colddiffusion after which the sample can be safely removed and submitted toa heated formalin step for crosslinking: duplicate capture ofhandwritten Lab Requisition Form by means of Electronic Data Capture andassociating it to a specific Specimen or Cassette via Barcode/RFID,verifying minimum fixation conditions; additional data capture ofbarcodes via camera, or radio frequency identifier (RFID)/near fieldcommunication (NFC) tags via built in communication interface;additional capture of an image of surgical site using webcam, etc.;and/or enabling such data capture for a Clinical Trials, in one exampleembodiment.

Cooling Box for Specimen Transport

FIG. 19 is an example diagram 6000 of an example cooling box forspecimen transport including cooling elements pre-charged over night fornext day surgery, reagent containers/vials kept precooled in a fridge, abox can be configured prior to surgery and transported to surgerylocation and can keep the reagents cold, and shipping can be initiatedat end of surgery day, according to one example embodiment.

Cooling elements can be pre-charged over night for next day surgery,according to one example embodiment.

Reagent Containers/Vials can be kept precooled in fridge, according toone example embodiment.

The box can be configured prior to surgery and transported to surgerylocation and can be configured to keep reagents cold, according to oneexample embodiment.

Shipping can be initiated at end of surgery day, or other appropriatetime, e.g., when a box is completely filled, according to one exampleembodiment.

The embodiments disclosed herein can be used with a range of differenttypes of fixatives, including standard tissue fixatives or non-standardtissue fixatives. Non-standard tissue fixatives in accordance with atleast some embodiments of the present technology can include formalinhaving a relatively high grade corresponding to a relatively highformaldehyde concentration. For example, the formalin can have a gradefrom about 16% to about 80%, from about 22% to about 80%, from about 16%to about 60%, from about 25% to about 60%, or within another suitablerange. Furthermore, a non-standard tissue fixative can include aphosphatase inhibitor compound, such as a phosphatase inhibitor compoundhaving a molecular weight less than about 300 Daltons. Suitablephosphatase inhibitor compounds include sodium fluoride, sodiumorthovanadate, sodium pyrophosphate, and beta-glycerophosphate, amongothers. In at least some embodiments, a tissue fixative includes morethan one phosphatase inhibitor compound. Non-standard tissue fixativesconfigured in accordance with at least some embodiments of the presenttechnology include a first phosphatase inhibitor compound, a secondphosphatase inhibitor compound, and, in some cases, a third phosphataseinhibitor compound, or more. The first, second, and third phosphataseinhibitor compounds can have molecular weights less than 300 Daltons andcan be selected to preferentially inhibit different phosphatases. Insome embodiments, the first, second, and third phosphatase inhibitorcompounds are selected from the group consisting of sodium fluoride,sodium orthovanadate, sodium pyrophosphate, and beta-glycerophosphate.In other embodiments, one or more of the first, second, and thirdphosphatase inhibitor compounds can be selected from other suitablegroups. These fixatives can be used during transport and/or during thefixation process.

At least some embodiments of the present disclosure monitor tissuesamples to improve preservation of biological molecules, tissuemorphology, and/or post-translational modification signals. Tissuesamples can be pre-soaked by submerging them in pre-cooled fixative. Thepre-soaking process can occur while the tissue samples are transportedto reduce overall processing times and to provide fast and accuratemedical diagnosis. The pre-soaking process can be monitored to obtaininformation about the pre-soaking process or other information used forsubsequent tissue processing, such as a histological processing (e.g.,fixing, embedding, dehydrating, infiltrating, embedding, sectioning,and/or staining). The information about the pre-soaking process can beused to determine, for example, (1) diffusion status of tissue samples,(2) target temperatures for fixation, and (3) tissue processing times(e.g., when to remove the tissue samples from fixatives). After fixingthe tissue sample, the tissue sample can be removed from the fixativeand can be subjected to one or more histological processes performedbased, at least in part, on information obtained from the monitoring.

To minimize or limit fixation during shipping of the tissue sample(e.g., shipping from a collection site to a processing site), thepre-cooled fixative can be at a pre-soaking temperature for a pre-soakperiod of time. The pre-soak process can involve diffusion of coldfixative throughout substantially the entire thickness or cross sectionof the tissue sample. After desired diffusion is achieved, the tissuesample can be heated to a fixation temperature that is higher than thepre-soak temperature to start and/or promote cross-linking. Byinhibiting or preventing fixation during transport, the fixation processcan be performed at the processing site to accurately control thefixation process. If the fixative is formaldehyde, cross-linking canoccur between formaldehyde molecules and amine containing cellularmolecules without significantly compromising the tissue characteristics(e.g., antigenicity, morphology, or both).

FIG. 20A is a workflow for processing a tissue sample in accordance withone embodiment. Generally, workflow system 100 is used to monitor tissuesamples from sample collection through processing. Collected informationcan be used to perform subsequent histology processing and to generatereports (e.g., reports used for diagnosis, patient monitoring, billing,etc.), an audit trail (e.g., an audit trail of specimen handling steps),a processing parameter log (e.g., a log that could be printed and as aquality record at the end of the processing), or the like.

At 102, a tissue sample is taken from a subject and can be a section oftissue, an organ, a tumor section, a smear, a frozen section, a cytologyprep, or cell lines. An incisional biopsy, a core biopsy, an excisionalbiopsy, a needle aspiration biopsy, a core needle biopsy, a stereotacticbiopsy, an open biopsy, or a surgical biopsy can be used to obtain thesample. In some embodiments, the tissue sample can contain genomic DNA,RNA (including mRNA), protein, or combinations thereof. Example tissuesamples include, but are not limited to, peripheral blood, urine,saliva, tissue biopsy, surgical specimen, amniocentesis samples andautopsy material. In one example, a sample includes a biopsy of anadenocarcinoma, a sample of noncancerous tissue, and a sample of normaltissue (from a subject not afflicted with a known disease or disorder).

At 106, the tissue sample is placed in a transporter system with media.A freshly removed tissue sample can be pre-soaked by placing the tissuesample in fixative within an appropriate amount of time to prevent orlimit an appreciable amount of degradation (e.g., ischemia). In someembodiments, the tissue sample is taken from a subject and placed in thefixative within a relatively short amount of time, for example, lessthan about 1 minute, 2 minutes, 5 minutes, 30 minutes, 1 hour, 2 hours,or the like. To pre-soak the tissue sample, the tissue sample can beplaced into an open transport specimen container containing liquidfixative. The composition and temperature of the fixative can beselected to achieve a desired rate of diffusion and/or cross-linking.The fixative can be formalin solutions of formaldehyde in water used forpreservation of biological specimens. Formalin used as a fixativetypically is 10% NBF, but other solution concentrations (e.g., 10%-80%NBF) also can be used. The tissue sample can remain immersed in thefixative while it is conveniently transported to another location.

Exemplary transporter systems can include, but are not limited to,packaging, a bottle, a vial, or other object used to hold liquid mediaand at least one tissue sample. The transporter systems can includemachine-readable code (e.g., optical symbology, magnetic pattern orelectromagnetic, or electrostatic signal having information content)that may relate to sample identity, patient information, sample origin,sample chain of custody, instructions for processing samples,information regarding the characteristics of samples, test results forsamples, images of samples, or other information associated with thetissue sample.

The fixative can be at a pre-soak temperature (or “diffusiontemperature”) to allow diffusion of the fixative while inhibitingfixation. The pre-soak temperature can be above the freezing point ofthe fixative solution to about 15° C., preferably greater than 0° C. toan upper temperature to about 10° C., more preferably greater than 0° C.to an upper temperature of about 7° C., and even more preferably fromabout 3° C. to about 5° C. For some procedures, the pre-soak temperaturecan be about 4° C. Although some fixing (e.g., cross-linking) may occurduring the pre-soaking process, the fixing primarily occurs after thepre-soaking process (i.e., during the fixation process). The pre-soakingprocess can balance the beneficial properties associated withsubstantially complete diffusion while minimizing or limiting theeffects associated with initializing or promoting cross-linking. In someembodiments, the rate of diffusion of the fixative can be maximizedwhile limiting and minimizing any deleterious effects associated withincreased cross-linking rate. In other embodiments, the transportersystem can include warm fixative (e.g., room temperature) thatsimultaneously diffuses through the tissue sample and cross-links.

At 108, the transporter system can be transported to an onsitelaboratory or an offsite site laboratory while the fixative diffusesthrough the tissue sample. The fixative can be kept at or below thepre-soak temperature, thereby allowing for a wide range of deliverytimes without significantly impacting the state of the tissue specimen.In some procedures, the fixative is formalin with a target pre-soakperiod of time in a range of about 15 minutes up to about 4 hours, mosttypically from greater than 15 minutes to about 3 hours, with desirableresults typically being obtained by immersing tissue samples for about1.5 hours to about 2 hours. Tissue samples with different sizes andcharacteristics can have different target pre-soak periods of time.Increasing the pre-soak period of time to 4 hours or greater may havelittle beneficial effect for a relatively small tissue sample with athickness up to 4 mm. If the sample arrives at the processing sitebefore completion of the target pre-soak period of time, the tissuesample can be left in the fixative to complete the diffusion process.Alternatively, the tissue specimens can be removed from the transportersystem and the pre-soaking process can be completed with anotherfixative.

At 110, the tissue sample can be evaluated to determine whether thefixative has adequately infused the sample tissue. After desireddiffusion is achieved, the fixation process can be performed by, forexample, heating the tissue sample to a fixation temperature to startand/or promote cross-linking if the rate of cross-linking is temperaturedependent. In some procedures, the tissue sample can be removed from thetransporter system and delivered to a processing apparatus/system thatcan perform a fixation process by immersing the tissue sample in warmfixative. If the fixative is formaldehyde, the temperature of the warmformalin can be greater than the ambient temperature and up to at least55° C., more typically from about 35° C. to about 45° C., as thistemperature range may increase the cross-linking kinetics sufficientlyto allow relatively quick tissue cross-linking. However, if thetemperature is increased above about 50° C., the tissue sample may beginto degrade, which may have a deleterious effect on certain subsequenthistological reactions. Thus, the upper temperature and time period ofthe fixation process can be selected to allow subsequent imagingprocesses, such as in situ hybridization, IHC, and/or hematoxylin andeosion (H&E) processing. The time period for the fixation process canrange from greater than about 15 minutes up to at least about 5 hours,more typically is at least about 1 hour to about 4 hours, and moretypically is from about 2 hours to about 3 hours. In certainembodiments, the fixation process can be performed for about 1.5 hour ata temperature of about 45° C. The pre-soak fixative and thecross-linking fixative can be the same or different. As yet anotherexample, entirely different aldehyde fixatives, such as formaldehyde andglutaraldehyde, can be used for the pre-soak process and fixationprocesses. Additionally, instead of removing the sample from thefixative, the pre-soak fixative can be heated to cause cross-linking. Assuch, the tissue sample can remain immersed in the same fixativethroughout the pre-soak and fixation processes.

Various factors may be considered to determine pre-soaking and fixationprocessing. These factors can include: sample thickness, which typicallyranges from about 1 mm to about 10 mm thick, more typically from about 2mm to about 8 mm thick, and even more typically from about 4 mm to about6 mm thick; volume of fixative to tissue sample mass, which typically isfrom about 10:1 to about 50:1 volume to mass; fixative composition;temperature; and sample immersion time in the fixative. Processingtimes, fixation history, condition history, tissue characteristics, orother histology information can be used to adjust processing to ensureproper tissue processing.

At 114, the tissue sample can be embedded, sectioned, and transferredonto a microscope slide for subsequent processing and analyses, such asstaining, IHC, or in situ hybridization. To section a tissue sample foroptical microscope examination, a relatively thin strip of tissue can becut from a large tissue sample so that light may be transmitted throughthe thin strip of tissue. A microtome can cut the specimen into thinsections, for example, slices on the order of about 5 microns to about 6microns thick. Each section can include a portion of the tissue sampleand some of the embedding material. The microtome and any otherequipment (e.g., a staining station, an embedding station, an oven,etc.) can include communication devices to read and/or write informationto the specimen holder. The tissue specimen can be transferred onto amicroscope slide, which can include machine-readable code. In someembodiments, the cut sections are floated on water to spread or flattenthe sections. If the sections are pieces of paraffin embedded tissue,the sections can be floated on a warm bath to keep the sections ingenerally flat configurations, thereby reducing or preventing folding,creasing, or bending. A microscope slide is inserted into the warm bath.A front surface of the slide is used to pick up the tissue specimens. Toexamine multiple tissue samples (e.g., a set of tissue samples, eachtaken at a different location in a subject) using a single slide, aplurality of the tissue samples may be sequentially floated onto theslide. These wet slides are then dried using the slide dryer andcoverslipped. Other standard tissue processing protocols can be used toanalyze the tissue samples.

FIG. 20B is a workflow 190 for tracking tissue samples from a collectionsite to a processing site. At 200, a tissue sample can be taken from asubject at an operating room and loaded into a specimen container. Thespecimen container can be assigned identification information and loadedinto shipping container, such as a transporter system. At 204, thetissue sample is shipped from the collection site to the processing site(such as an onsite or offsite pathology lab) by, for example, a courieror a runner. The transport time can be less than 24 hours. At 206,information (e.g., temperature condition, temperature versus timeinformation, diffusion state, etc.) from a monitoring system of thetransporter system can be used to determine an appropriate fixationprotocol. The tissue sample can be fixed, and the fixed tissue samplecan be processed using standard tissue processing techniques.

FIG. 20C is a front view of a self-cooling transporter system 200(“transporter system 200”) in accordance with one embodiment. Thetransporter system 200 includes a transport or specimen container 210(“transport container 210”), a cooling device 230, and a monitoringsystem 232. The cooling device 230 can adjust the temperature of thecontents of the transport container 210. The monitoring system 232 canobtain and store information about the contents of the transportcontainer 210. The stored information can be used to determine thecondition of tissue sample(s) carried by the transport container 210.

FIG. 20D is a cutaway view of the transporter system 200 of FIG. 20C.The transport container 210 has a holding chamber 212 containing mediain the form of fixative 214 and a tissue sample 220. The fixative 214can be any of the fixatives disclosed herein. The cooling device 230 isoperable to reduce a temperature of the fixative 214 before, during, orafter placing a tissue sample 220 into the container 210. In someembodiments, the cooling device 230 can be a fixation-inhibiting coolingdevice that reduces the temperature of the fixative 214 from roomtemperature (e.g., about 20° C.-about 25° C.) to a pre-soak temperature(e.g., temperature less than about 5° C.) before placing the tissuesample into the transport container 210. The pre-soak temperature can besufficiently low to inhibit fixation of the tissue sample 220 and can beselected based on the characteristics of the fixative. In someembodiments, the temperature of the fixative 214 can be reduced to atemperature equal to or lower than the pre-soak temperature and/ormaintain the pre-cooled state. For example, the cooling device 230 canreduce an average temperature of about 400 ml, 200 ml, or 100 ml of thefixative 214 at least about 25° C. in less than about 5 minutes, 4minutes, or 3 minutes. In some embodiments, the cooling device 230reduces a temperature of the fixative 214 at least about 25° C. in lessthan about 5 minutes and can periodically cool the fixative 214 asneeded.

In other embodiments, the transport container 210 can be configured tohold relative large specimens, such as resected material, partialorgans, whole organs, or the like Such containers 210 can hold one ormore liters of fixative to perform relative long pro-soak procedures of,for example, 6 hours, 1 day, or multiple days, depending on the size ofthe specimen(s). In some embodiments, transport container 210 isconfigure to reduce an average temperature of at least one 1 liter offixative in the specimen container by at least 15°. This cooling can beperformed in less than 1 hour, 30 minutes, 10 minutes, or 5 minutes. Theholding capacity and cooling capacity of the transport container 210 canbe selected based on the size of the specimen, length of pre-soakprocedure, and type of fixative.

To cool the fixative 214, the cooling device 230 can be changed from anon-cooling or standby mode to a self-cooling cooling mode to reduce thetemperature of the fixative 214 without utilizing external power sources(e.g., electrical outlets), refrigerators, freezers, or the like. Thefixative 214 can also be cooled during transport. As such, thetransporter system 200 can be conveniently used at a wide range oflocations.

The transport container 210 can include a main body 240 and a removableclosure 250. The main body 240 can be thermally insulated to inhibitheat transfer between the surrounding environment and the fixative 214.The wall thickness t, thermal characteristics (e.g., thermalconductivity), holding capacity (e.g., 50 ml, 100 ml, 200 ml, 400 ml,etc.), and configuration of the transport container 210 can be selectedbased on the pre-soak process to be performed. In some embodiments, thetransport container 210 can keep the fixative 214 at or below about 10°C. for at least one hour while an ambient temperature surrounding thetransport container equal to or higher than 10° C. In one embodiment,the fixative 214 can be kept at less than about 4° C. for at least about2 hours while the ambient temperature is about 25° C.

The transport container 210 can also be disposable. As used herein, theterm “disposable” when applied to a component (or combination ofcomponents), such as a container or cassette, is a broad term and means,without limitation, that the component in question is used a finitenumber of times and then discarded. Some disposable components are usedonly once and then discarded. Other disposable components are used morethan once and then discarded. For example, the transport container 210can be a single-use container made, in whole or in part, of a relativelyinexpensive material suitable for contacting fixative.

The transport container 210 has an open configuration (FIG. 20D) forloading/unloading tissue samples and a closed configuration (FIG. 20C)for sealing the holding chamber 212. The closure 250 can be coupled tothe main body 240 to form a seal (e.g., a hermetically seal, afluidically seal, etc.) to prevent leaking of the fixative 214. In oneembodiment, the closure 250 can be a screw cap. In other embodiments,the closure 250 can be a flip cap movable between an open position foraccessing the chamber 212 and a closed position for closing the chamber212.

FIG. 20D shows the cooling device 230 in thermal communication with thefixative 214 and including a first chamber 300, a second chamber 310,and a mixing chamber 312. Substances from the chambers 300, 310 can mixin the mixing chamber 312 in response to the cooling device 230 changingfrom a non-cooling mode to a cooling mode by depressing a button or anactuator 320. The substances can include, without limitation, agentsthat react to produce an endothermic reaction or phase change materials.In some embodiments, the chamber 300 holds an activation agent in theform of water and the chamber 310 holds a cooling agent. The coolingagent can include, without limitation, a nitrogen compound, a potassiumcompound, or a phosphorus compound. Additionally or alternatively, thecooling device 230 can include, without limitation, one or more phasechange materials that undergo a phase change when heated so as to coolthe fixative 214. For example, the chamber 300 can store a desiccant andthe chamber 310 can store a substance that evaporates to cool thefixative 214. In other embodiments, the cooling device 230 can include,without limitation, a refrigeration unit, a thermoelectric cooler unit,or like. The refrigeration unit can operate on a vapor-compressioncycle, a vapor-absorption cycle, and the like. Thermoelectric coolerunits can include, without limitation, one or more power sources (e.g.,batteries) and Peltier devices that operate to cool, or heat, thefixative 214. In other embodiments, the cooling device 230 can allowexpansion of gas to cool the fixative. For example, a fluid (e.g., CO₂gas) can pass through a valve opened by a user to change the coolingdevice 230 from a non-cooling mode to a cooling mode. The fluid can flowthrough the opened valve and expand to absorb heat, thereby reducing thetemperature of the fixative 214. The cooling capacity, configuration,and modes of operation of the cooling device 230 can be selected basedon the volume of the fixative 214, pre-soak period of time, predictedambient temperature range, and/or fixative properties.

Referring to FIGS. 20C and 20D, the monitoring system 232 can include asensing element 330 positioned in the chamber 212, within a wall of themain body 240, on an outer surface 342 of the main body 240, or atanother suitable location for detecting the temperature of the fixative214. The sensing element 330 can include, without limitation, one ormore thermistors, thermocouples, or other temperature sensors configuredto periodically or continuously measure temperatures over a period oftime. In some embodiments, the monitoring system 232 can further includea temperature data logger to store such measurements and/or atime-temperature indicator device capable of providing a cumulativetime-temperature history. In some embodiments, one time-temperatureindicator device measures the cumulative time-temperature history of thefixative 214 and another time-temperature indicator device measures thetime-temperature history of the ambient environment. Exemplarytime-temperature indicator devices can provide visual indications of thetime history and may include, without limitation, a timer (e.g., adigital timer), a color changing temperature strip, a timer and/ortemperature gauge based on dye diffusion through an absorbent strip, atemperature sensor, or combinations thereof, as well as memory,transmitters, receivers, and other storage or communication elements.

FIG. 20E is a cross-sectional view of the transporter system 400 of FIG.2. A cooling device 470 is positioned between the tray 436 and a bottom474 of the main body 410. The cooling device 470 can include athermoelectric unit, a cooling plate, or other component capable ofactively removing heat from the tray 436, as well as the containers 422.The communication device 452 can provide power via a power line 476.Other types of cooling devices can be used to cool or heat the specimencontainers 422.

The controller 450 can include input devices (e.g., a touch pad, a touchscreen, a keyboard, or the like), peripheral devices, memory,controllers, processors or processing units, or combinations thereof. Tostore information, the comptroller 450 can also include memory that caninclude, without limitation, volatile memory, non-volatile memory,read-only memory (ROM), random access memory (RAM), and the like. Theinformation can include, but is not limited to, patient information,measurements from the detection devices 442, protocols (includingmonitoring protocols, tracking protocols, etc.), data (includingdatabases, libraries, tables, algorithms, records, audit trails,reports, etc.), settings, or the like. The measurements can includetemperature measurements indicative of the temperature of tissuespecimens. Such measurements can include the temperature of the walls ofthe specimen containers 422 or temperature of the fixative therein. Whenthe transporter system 400 enters the laboratory, the controller 450 cancommunication information to processing equipment (e.g., tissueprocessing system) at the laboratory via a network, such as a wirelessnetwork at a pathology lab.

In some embodiments, the detectors devices 442 can be TOF sensors formeasuring changes in the TOF of sound in tissue samples by, for example,comparing the acoustic waves outputted by the transmitter to theacoustic waves detected by the receiver. This comparison can be repeatedany number of times to monitor the fixation state of the tissue samples.In some embodiments, the controller 450 determines a first length oftime it takes the acoustic waves to travel through the tissue samples.The controller 450 can then determine a second length of time it takes asubsequently emitted acoustic wave to travel through the tissue samples.The first length of time is compared to the second length of time todetermine, without limitation, a change in speed (e.g., acceleration) ofthe sound waves, an absolute and/or relative change in TOF, orcombinations thereof. The TOF measurements can be used to monitor thediffusion process throughout transport.

Although the cooling device 470 can be configured to cool or heat thespecimen containers 422 (and their contents), the specimen containers422 can also include cooling devices. FIG. 20F shows a cooling device492 positioned in a chamber 494. The cooling device 492 can be similarto the other cooling devices discuss herein, such as the cooling device230. The cooling device 492 can reduce the temperature of a fixative 498before, during, or after loading the tissue sample 493. In someembodiments, about 100 ml of fixative 498 can be cooled from an elevatedtemperature (e.g., room temperature of 22° C.-25° C.) to a loweredtemperature (e.g., 4° C.) immediately prior to loading of the tissuesample 493. The cooling device 492 can continuously or periodically coolthe fixative 498. During transport, the cooling device 492 can helpreduce the temperature of the specimen container 422 to reduce thermalloading of the cooling device 492.

FIG. 20F shows the specimen container 422 having a closure 502 with aholder 504 for keeping the tissue sample 493 from contacting a wall 413and/or bottom 415. In some embodiments, the holder 504 can be a clip. Inother embodiments, the holder 504 can be a disposable or reusablecassette coupled to a cap.

FIG. 20G shows a transporter system 500 in accordance with anotherembodiment. The transporter system 500 includes a transport container502, a cooling element 510, and specimen containers 520 (individually520 a, 520 b, 520 c, 520 d). The containers 502 can be insulated potspre-filled with fixative and kept in a refrigerator. One example of sucha container can be CELLSTOR POT available from CellPath Ltd. (Newtown,Powys, UK). Other types of containers can also be used. The coolingelement 510 can be in the form of a heat sink and can be stored in afreezer at a relatively low temperature (e.g., at a temperature lessthan about −20° C.) but in other embodiments can be a fixationinhibiting cooling device (including self-cooling devices). Thetransport container 502 can include a closure 511 (e.g., a cover or alid) and a thermally insulated main body 513. The cooling element 510can include a datalogger 526 and/or a temperature sensor 527 (e.g.,temperature strip) to confirm that the cooling element 510 is at adesired temperature. To ship the sample, the cooling element 510 can betaken from the freezer and placed into a chamber 523. The temperature ofthe cooling element 510 can equilibrate to about 2° C. to about 4° C.The samples can be placed into the specimen containers 520 pre-filledwith fixative and loaded into the main body 513. After loading thepatient samples into the individual containers 502, a timer 532 can beactivated to track (e.g., to track transport/processing time informalin) each sample individually. The main body 513 can contain iceand/or chilled liquid to keep the containers 502 at a low temperature.The closure 511 can then be placed on the placed on the main body 513.The packed system 500 is then ready to be shipped or transported to aprocessing site.

Upon arrival at the processing site, the temperature strip 527 and/ortemperature sensors 531 (individually 531 a, 531 b, 531 c, 531 d) can bechecked to confirm that the fixative and/or tissue samples are within adesired temperature range (e.g., 2° C. to about 8° C.) and, in someembodiments, can be time-temperature strips that can be inspected toverify progression of dye (e.g., a blue dye) of at least 2 hours, butnot more than 24 hours. The timer 532 can indicate the length ofpre-soaking and/or shipping time. If additional cold time is needed toreach a target pre-soak period (e.g., 2 hours), the containers 502 canbe returned to the container 502 or kept in a chiller (e.g., arefrigeration unit) at a temperature of about 4° C. After the fixativehas diffused through the tissue samples, the tissue samples can betransferred to a histology cassette and a tissue processor to perform afixation process. In some fixative procedures, tissue samples areprocessed for about 2 hours in a fixative (e.g., 10% NBF-40% NBF) atabout 45° C. If needed, the tissue samples can be held in alcohol (e.g.a 70% alcohol solution) for extended periods of time.

FIG. 20H shows a monitoring system 600 for analyzing tissue sample 640.FIG. 20I shows the container 639 with a chamber 680 filled with aprocessing media 670. The processing system 600 includes a specimenholder 610, a container 639 filled with media 670, and an analyzer 614positioned in the container 641. The analyzer 614 includes a transmitter620 and a receiver 630. A computing device or controller 660 iscommunicatively coupled to the analyzer 614. The specimen holder 610,the transmitter 620, and the receiver 630 are submerged in theprocessing media 670. A thermal device 662 (FIG. 20I) can increase ordecrease the temperature of the media 670 to, for example, performprocesses at different temperatures. The thermal device 662 can include,without limitation, one or more refrigeration devices, heaters (e.g.,resistance heaters, electric heaters, etc.), thermoelectric devices(e.g., Peltier devices), or the like. Additionally or alternatively, adevice 663 (FIG. 20I) can include a mixing element (e.g., a mixing rod,a mixer, etc.) for mixing or agitating the media 670.

To pre-soak a tissue sample 650 (e.g., to complete a partially completedpre-soak process), the media 670 can be a cold fixative at a temperatureof about 0° C. to about 5° C. The computing device 660 can cause thetransmitter 620 to output energy that passes through the tissue sample650. The receiver 630 can receive the energy and can send signals to thecomputing device 660 in response to the received energy. The computingdevice 660 analyzes those signals to monitor processing. After thedesired level of diffusion is achieved, the thermal device 662 canincrease the average temperature of the media 670 to promotecross-linking. Once fixation is achieved, the sample 650 can be removedfrom the media 670. In other embodiments, the processing system 600 isused to only perform a fixation process.

In some embodiments, the transmitter 620 can output acoustic waves(e.g., infrasound waves, audible sound waves, ultrasound waves, orcombinations thereof) such that propagation of the acoustic wavesthrough the specimen 650 change because of changes to the tissue sample650. During diffusion, the acoustic properties of the tissue sample 650can change as the media 670 infuses the tissue sample 650. Duringcross-linking, mechanical properties (e.g., an elastic modulus) of thetissue sample 650 may change significantly as cross-linking progressesthrough the tissue. The change in elastic modulus may alter the acousticcharacteristics of the tissue sample 650. Acoustic characteristicsinclude, without limitation, sound speeds, transmission characteristics,reflectance characteristics, absorption characteristics, attenuationcharacteristics, or the like. To evaluate transmission characteristics,a time of flight of sound (e.g., audible sound, ultrasound, or both),the speed of sound, or the like can be measured. The TOF is a lengthtime that it takes for acoustic waves to travel a distance through anobject or substance. In some embodiments, the TOF is the length of timeit takes acoustic waves to travel through a specimen in comparison tothe time to travel through the medium displaced by the specimen. In someembodiments, the time of flight of the medium and the measurement device(e.g., the holder) may be recorded prior to insertion of the sample andstored for later reference so that it can be used for temperaturecompensation, evaporative losses, compensation protocols, predictivemodeling, or the like. The thickness of the specimen 650 can besufficiently large to produce a measurable change in the TOF. Inreflectance embodiments, the TOF can be the length of the time theacoustic waves travel through a portion of the tissue sample. Forexample, the TOF may be the length of time that the acoustic wavespropagate within a portion of the tissue sample. Thus, the TOF can becalculated based on acoustic waves that travel through the entirespecimen, acoustic waves reflected by the tissue sample, or both.

The computing device 660 can evaluate the change in the TOF of sound inthe tissue sample 650 by, for example, comparing the acoustic wavesoutputted by the transmitter 620 to the acoustic waves detected by thereceiver 630. This comparison can be repeated any number of times tomonitor the state of the tissue sample 650. In some embodiments, thecomputing device 660 determines a first length of time it takes theacoustic waves to travel through the tissue sample 650. The computingdevice 660 then determines a second length of time it takes asubsequently emitted acoustic wave to travel through the tissue sample650. The first length of time is compared to the second length of timeto determine, without limitation, a change in speed (e.g., acceleration)of the sound waves, an absolute and/or relative change in TOF, change indistance between the transmitter 620 and the receiver 630, change intemperature and/or density of the processing media 670, or combinationsthereof. The computing device 660 can use different types of analyses,including a phase shift analysis, an acoustic wave comparison analysis,or other types of numerical analyses.

To store information, computing device 660 can also include memory.Memory can include, without limitation, volatile memory, non-volatilememory, read-only memory (ROM), random access memory (RAM), and thelike. The information includes, but is not limited to, protocols, data(including databases, libraries, tables, algorithms, records, audittrails, reports, etc.), settings, compensation schemes, or the like.Protocols include, but are not limited to, baking protocols, diffusionprotocols, fixation protocols, tissue preparation protocols, stainingprotocols, conditioning protocols, deparaffinization protocols,dehydration protocols, calibration protocols, frequency adjustmentprotocols, decalcification protocols, or other types of routines.Protocols that alter or impact tissue density or sound transmission canbe used to control the components of the computing device 160,components of the analyzer 614, microscope slide processing units,stainers, ovens/dryers, or the like. Data can be collected or generatedby analyzing the specimen holder 610, the processing media 670, thetissue sample 650, or it can be inputted by the user.

FIG. 20J shows a monitoring system 700 that includes an analyzer 702with a reference sensor 710 and a specimen sensor 712. The analyzer 702can be moved to position one of the sections or portions 720(individually 720 a-f) between a transmitter 730 and receiver 732 of thespecimen sensor 712, and the reference sensor 710 includes a transmitter750 and a receiver 752. The computing device 660 can evaluate thecharacteristics of the portion 720 based on the output from the sensors710, 712 and can compensate for temperature fluctuations.

FIG. 20K shows an analyzer 800 including a carrier assembly 802 with anacoustic sensor assembly 804. The sensor assembly 804 can include atransmitter unit 821 and a receiver unit 823. The transmitter unit 821can include transmitters 822 (with one transmitter labeled 822). Thereceiver unit 823 can include receivers 824 (with one transmittedlabeled 824) that receive acoustic waves from respective transmitters822. The pattern, position, and spacing of the transmitters 822 andreceivers 824 can be selected based on the tissue specimens to beanalyzed. FIG. 20L shows the carrier assembly 802 further including aspecimen holder 851 positioned to hold a tissue sample at a detectionzone 861. In some embodiments, the specimen holder 851 can hold acassette or other type of holder.

FIG. 21A is an isometric view of a tissue processing apparatus 900including containers (one container is labeled 922) holding reagents. Atransport mechanism 912 can move the carrier assembly 802, as indicatedby arrow 916, to submerge the sensor assembly 804 and tissue sample inreagent baths. The transport mechanism 912 can include, withoutlimitation, a 3-axis handling robot or other type of robotic equipment.A carriage 919 can be moved along a rail 927 to lower the carrierassembly 802, such that a cover 817 closes the container 922. A sealingmember can be used to form a seal between the cover 817 and thecontainer 922. A shaft 925 can be moved to move the specimen holder 851relative to the sensor assembly 804. For example, the specimen holder851 can be moved from a first position (shown in FIG. 20L) to a loweredposition (shown in phantom line in FIG. 20L). After the carrier assembly802 is moved from a raised position to a lowered position, the sensorassembly 804 can analyze the submerged tissue specimen and sequentiallyscan multiple positions in the vertical direction, thus allowing to thesensor assembly 804 to obtain a 2-dimensional image or impression of thesample and its related TOF profile. A computing device 952 iscommunicatively coupled to the sensor assembly 804 and can evaluate thespecimen based on time of flight changes. In some embodiments, includingthe illustrated embodiment, one container 922 can hold a fixative, andother containers can include other processing media, such as clearingagents (e.g., xyline or the like), infiltrations, dehydration agents, orreagents. After fixing, the specimens can be sequentially delivered toother containers.

FIG. 21B is a plot of frequency versus phase comparison in accordancewith one embodiment. A programmable function generator can generate manysine waves at different frequencies. By measuring the phase differenceat each frequency, we can reconstruct the actual time-of-flight. Acomparison can be performed as a function of frequency to unique time offlights. As shown in FIG. 21B, the comparison wave can be a generallytriangle wave with a slope corresponding to the time-of-flight. TOF canalso be determined based on techniques disclosed in U.S. applicationSer. No. 13/372,040, filed on Feb. 13, 2012, and incorporated byreference in its entirety.

FIG. 21C is a plot of temperature versus time of flight in media. Thecurve shows the media time of flight gradually decreasing as thetemperature of the media increases. As such, temperature changes cancause significant changes to time of fight. In some fixation medias,every degree (e.g., every ° C.) of change can change the target signalby about 30 nanoseconds in a 25 mm acoustic path. The 25 mm acousticpath can include 20 mm of media (i.e., the distance through the media)and 5 mm through the tissue sample. Because the length of the acousticpath is longer in the media (i.e., the media path of 20 mm is greaterthan the 5 mm thickness of the tissue sample), the changes in themedia's time of flight can be minimized or limited by controlling thetemperature of the media. In some embodiments, the temperature of themedia can be held at a generally constant temperature to minimize orlimit changes in time of flight caused by temperature changes.

Different schemes can be used to compensate for environmentalconditions. FIG. 21D shows a tissue sample time of flight curve 1002 anda media temperature curve 1004. In one compensation scheme, a referencepath time of flight readout is used in a control scheme. For example, acontroller can operate on a proportional-integral-derivative (PID)control loop using a set value 1010 of the total time of flight. The setvalue 1010 can be close to the target temperature. The controller canalso apply a proportional term correction to achieve temperature controlof the media on the order of 1/100° C. relative precision. This canresult in time of flight variation of the media of less than 1nanosecond, thereby allowing better interpretation of the true time offlight of the target tissue. Other compensation schemes can also be usedto compensate for temperature changes, as well as other environmentalconditions (e.g., temperature fluctuations during shipping).

FIG. 21E is a plot of time versus a reference time of flight. Curve 1012is a warmup curve using a reference time of flight for temperaturecompensation. The reference time of flight gradually decreases untilreaching a generally constant time of flight 1014 at a steady statetemperature.

FIG. 21F is a plot of time versus a target time of flight. The curve isa thermal stability curve for a target time of flight of a targetsample. The curve can be obtained using the analyzers, systems, andtechniques disclosed herein. The general shape of the curve of FIG. 21Fis driven by the media TOF changes associated with temperature changes.

FIG. 21G is a plot of time versus the target time of flight using thecurve 1012 (FIG. 21E) to compensate for temperature changes. The systemsdisclosed herein can include different data sets, curves, curve fittingalgorithms, and compensation algorithms or schemes to minimize or limitchanges in time of flight caused by temperature changes of the media.The curve of FIG. 21G corresponds to the time of flight of the targetsample where the temperature contribution is eliminated. As such,temperature changes associated media (e.g., temperature changesassociated with transport or shipping) can be eliminated. The targetsignal to noise ratio can be increased by applying various methods, suchas averaging, filtering, Fourier analysis, and/or elimination ofperiodic signals. Other techniques can be used to enhance signaldetection.

Exemplary Two-Temperature Formalin Fixation Methodology

Two-temperature formalin fixation has been identified as a novelsolution to the problem posed by traditional formalin fixationmethodologies. Longitudinal research has indicated that heat-assistedformalin fixation, which dramatically reduces fixation time and hasbecome the gold-standard of laboratories worldwide, can compromise thequality of histology samples, resulting in downstream diagnostic errors,especially in biomarker-based applications (including phosphoproteinassays). In addition, as the field of companion diagnostics (whichnecessitates multi-site, multi-institutional cooperation) expands,continued variation in fixation protocols from lab to lab has beenidentified as a significant impediment to accurate pathologicalanalysis, and to the development of properly targeted diagnostics andtherapies.

Ventana Medical Systems, Inc. has developed a two-temperature, rapidformalin fixation protocol that reduces fixation time and is optimizedfor use across a broad range of tissue types through better preservationof histomolecular features, according to an exemplary embodiment. Thetwo-temperature process involves soaking the tissue specimen inprecooled formalin for up to five minutes and then letting the samplesoak for two hours at 4° C., and another two hours at 45° C., accordingto an exemplary embodiment. This can be generally performed on standardlaboratory equipment such as an ice bucket or heating plate, in anexemplary embodiment. Finally, dehydration, clearing, paraffinization,embedding and sectioning are generally performed in an automaticstandalone tissue processor (such as, e.g., but not limited to, Leica,Sakura, Avantik, etc.), according to an exemplary embodiment.

Exemplary Sample Collection, Transport and Tracking Solution

Ventana Medical Systems Inc. has devised an integrated specimencollection and transport solution that ensures specimens are optimizedfor subsequent staining and analysis using a cold packing method thatcan include the first half of the two-temp fixation process, accordingto an exemplary embodiment. By the time the pack is opened at theanatomical pathology (destination) laboratory, the specimen has beenproperly fixed at the appropriate cold temperature, according to anexemplary embodiment. The specimen can then be heated up, using, e.g.,but not limited to, standard laboratory equipment or specialized Ventanahardware, etc., for the requisite two hours, and can be subsequentlyready for tissue processing, according to an exemplary embodiment.

The integrated system can also provide digital tracking, in the form ofspecimen temperature and time recording that can enable pathologyprofessionals to monitor conditions that may affect subsequentprocessing or analysis, according to an exemplary embodiment. The systemis designed for use with either paper-based laboratory request forms, ora digital e-LRF pen that accompanies the specimen throughout its journeyand allows for paperless routing, as well as enhanced specimen trackingthat can be synced to wireless devices or uploaded for cloud computing,according to an exemplary embodiment.

Methodological and Logistical Problems of Conventional Transport andTracking

Some of the problems with conventional best-practice histology sample,collection, and transport methodologies include the following:

Current room-temperature fixation protocols do not preservephosphomarkers sufficiently.

Tissue storage after removal is not sufficiently standardized withrespect to phosphomarker preservation; this can result in some specimenseven being left without formalin over the weekend in a refrigerator. Thelack of rigid adherence to set protocols across institutions and sitesoften results in widely varying degrees of histomolecular degradation.This compromises downstream processing, especially in samples subjectedto a variety of biomarker-based diagnostics.

Current, standard specimen collection containers general contain asingle, initially-submerged tissue sample that does not stay in placeduring transport. This results in the tissue often floating up to thesurface, or adhered to the side of the wall of the container, thuscompromising the amount of contact each sample has with the fixativefluid, the perfusion of which is critical for proper preservation andsubsequent processing of the sample.

Fixative fluid spills as a result of inversion or jostling.

Variations in ambient temperature during transport, which has been knownto affect efficacy of fixative.

Operator exposure to caustic fumes during handling.

Exemplary problems with conventional standardized tracking methodologiesinclude:

General best-practices may vary intra- and inter-institutionally inspite of barcode labeling.

Current methodologies rely heavily on paper and individualized computerinput which may also vary from institution to institution; chain ofcustody is established through a series of signatures.

Real-time tracking of the computer specimen is paper-based and cannotprovide an up-to-the-minute accounting of temperature, location andtransport time.

The end result of these challenges is questionable sample quality(phosphomarker preservation may not correctly reflect the original stateof the tissue) and lack of traceability during processing. These areunacceptable risks when dealing with oncology patients.

An Exemplary Embodiment of the Solution

According to an exemplary embodiment, a sample collection, transport andtracking system is an integrated solution for the above-referencedchallenges. In an exemplary embodiment, the example sample collection,transport and tracking (SCTT) system can provide superior, digitizedpacking that can begin the process of optimized formalin fixationwithout supervision by a laboratory technician. The cold part of thefixation process can happen while the specimen is in transit, accordingto an exemplary embodiment. By beginning the cold part of the fixationprocess in transit, then once the specimens reach their destinationanatomic pathology laboratory, the specimens need only be heated therequisite 2 hours (at 45° C.) to achieve substantially higher quality(for biomarker-based diagnostics and treatment) fixation for subsequentprocessing, staining and analysis, according to an exemplary embodiment.

According to an exemplary embodiment, the container itself can offerjostle- (and potentially spill-, as discussed further herein in FIGS.28, 35, 37, 38, 39A and B, 40A-D, 41A-D, 42A and B, 43A-C, and 44A-E)proof, fully submerged tissue transport that can minimize operatorcontact and can ensure (e.g., but not limited to, through monitoring andfeedback alarms, etc.) steady thermal conditions. The entire process,according to an exemplary embodiment, can be recorded and/or monitored,with, e.g., but not limited to, real-time feedback to wifi-connectedtablet, and a digital pen that can establish a chain-of-custody fromsurgery/excision, through transport and handling, to laboratoryprocessing and subsequent analysis. In an exemplary embodiment, thedigital pen can be based on optical character recognition technology andcan accompany the tissue handling pod (containing an exemplary foursamples, in an exemplary embodiment, as illustrated in FIG. 3), whichcan enable surgeons, technicians, transporters, pathologists to pull upcomplete tissue data (e.g., but not limited to, tissue donor, testingprotocol, temperature and/or time log, etc.) at any point along thespecimen's journey, in an exemplary embodiment.

This integrated solution, according to an exemplary embodiment,therefore can result in, e.g., but not limited to, 1) superior formalinfixation results for a broader range of biomarker-based diagnostics,including, e.g., novel phosphomarker preservation previously onlypossible in fresh frozen specimens; 2) a processing time that iscompatible with standard fixation protocols (depending on tissue type);4) cold preservation of volatile markers for up to 72 hours; 4)error-free, fully electronic tracking establishing chain of custody thatcan be readily accessed from digital tablets and/or uploaded for cloudcomputing; 5) a unique tissue suspension system that keeps specimensfully, properly submerged with fluid fixative in spite of potentialinversion or jostling; 6) a real-time recording of thermal conditionsand time that can be used to gauge whether additional measures will benecessary at destination to ensure optimal processing, includingstaining.

Exemplary Kit Inventory

The specimen collection kit, according to an exemplary embodiment, canbe, and should be, assembled before going to the sampling site tocollect the specimen and should contain the following items: (See FIG.26 for an exemplary full cold tissue kit and FIG. 3 for an exemplarycold tissue assembly; and for full room temperature tissue kit andassembly, the assembly of FIG. 3, the depicted kit and assembly, exceptwithout cooling components 2206, 2208, 2214, and 2216)

3) 4 vials filled with 10% (alternatively, 30%) NBF;4) 4 total pods with Tempod dataloggers (alternatively, with cryopackdataloggers);5) 1 GreenBox with phase change blocks and vacuum insulated panels;6) 1 cardboard shipping box for each GreenBox;7) (Paper option) 4 Lab Request Forms (one for each Specimen); and8) (Electronic option) Digital pen and associated software (for allspecimens).

How the Exemplary Components Work Together:

FIG. 22 is an example process flow diagram 5200 illustrating an exampleworkflow variation between paper forms and e-LRF pen, illustratingexample information flow from pen to laboratory information system(LIS), according to an exemplary embodiment.

Flow diagram 5200 can begin with 5202, where during sample collection,the responsible party can fill out the eLRF, according to an exampleembodiment.

From 5202, flow diagram 5200 can continue with 5204, or 5206, accordingto example embodiments.

In 5204, the paper portion of the eLRF can travel with the tissuesamples to the AP lab, according to an example embodiment. From 5204,flow diagram 5200 can end.

In 5206, the digital pen can act like an USB stick with an electroniccopy of the eLRF data in memory, according to an example embodiment.From 5206, flow diagram 5200 can continue with 5208.

In 5208, the responsible party can upload the pen data from a PC, forexample, according to an example embodiment. When the data is extractedfrom the pen, the data can be transformed into a portable file formatsuch as, e.g., but not limited to, extensible markup language (XML),CSV, or XLS. From 5208, flow diagram 5200 can continue with 5210, 5214,or 5216.

In 5210, there can be several options as to where the responsible partycan upload the data, and information technology (IT) policy restrictionscan make it challenging to find a universal solution, according to anexample embodiment. From 5210, flow diagram 5200 can continue with 5212.

In 5212, the digital data from the pen may go directly to the APlaboratory in an electronic person-to-person communication like email,according to an example embodiment. The recipient may then load the datainto the AP Lab's LIS, according to one embodiment. From 5212, flowdiagram 5200 can continue with 5214, according to an example embodiment.

In 5214, the eLRF data can eventually arrive at the AP Lab's LIS,according to an example embodiment. If the LIS has internetconnectively, the LIS can send the eLRF data directly; or the eLRF datacan take one of the other paths to 5214, according to an exemplaryembodiment. From 5214, flow diagram 5200 can end, according to anexample embodiment.

In 5216, pen data can be uploaded to a server in the cloud, according toan example embodiment. From this uploaded data, approved parties canaccess the data. The AP Lab, the collection site, and the CRO (ifinvolved) can all access this data from a single source, according to anexemplary embodiment. From 5216, flow diagram 5200 can continue with5214, according to an example embodiment.

NBF vials (e.g., FIGS. 28, 35A-I, 37, 38, 39A and B, 40A-D, 41A-D, 42Aand B, 43A-C, and 44A-E)

FIG. 23A illustrates an exemplary sample vial 2302, according to anexemplary embodiment. The exemplary sample vials 2302 can be filled with10% neutral buffered formalin (NBF), according to one exemplaryembodiment. The exemplary specimen vials 2302 may also contain the 30%NBF for an exemplary further acceleration/optimization of the fixationprocess, according to one exemplary embodiment. The sample vials havemini-cassettes clamped to the underside of the specimen lid. This is themechanism whereby tissues are suspended upside-down in fluid. Thisorientation and cap design allow tissue to remain submerged in spite ofany vial or box manipulations/inversions.

FIG. 23A is an exemplary sample vial 2302, shown in illustration with anexemplary specimen lid 2402 affixed thereon using an exemplary threadedscrew coupling.

FIG. 23B depicts an example vial 2302 with exemplary screw threading,along with an example specimen lid 2402 (discussed further herein withreference to FIG. 4) with exemplary clip 3504 (discussed further belowwith reference to FIG. 35) coupled thereto, and an exemplary cassette2304 (discussed further below with reference to FIG. 23) clipped to thelid 2402 via the clip 3504, according to an exemplary embodiment.

FIG. 23C is another view of the cassette 2304 being inserted into theclip 3504 coupled to the specimen lid 2402, according to an exemplaryembodiment.

FIG. 23D depicts exemplary cassettes 2304, 2306 in an open position, anda closed position, according to an exemplary embodiment.

FIG. 24 is an example diagram 5500 of expected interactions processbefore processing, according to an exemplary embodiment. Diagram 5500illustrates external lab/collection site, of up to five, or more sites5502, may then be transported to an example centralized histology lab5304, in an example embodiment. Collection site 5502 can include, incombination 5506, a specimen collection management software and userinterface, as well as a prefilled container with integrated cassette2218. As noted, combination 5506, for 1-3 patients/day/site, processingcan be provided, or various data gathered 5520 (such as, e.g., but notlimited to, patient-ID, Study-ID, LabID, and for each specimen, load thespecimen into a cassette 2304, assign a cassette-ID, assign acontainer-ID to vial 2302 of pod 2404, etc.) and can be provided to amethod of transport 5524, according to an exemplary embodiment.Collection site 5502 can also include, in combination 5508, a coldstorage box 2200, as well as a loading monitor and a datalogger 2406,2408, in an example embodiment. As noted, from combination 5508,processing can be provided 5522, including, e.g., but not limited to,for each container or pod 2404, starting a datalogger 2406, 2408,assigning a datalogger-ID to vial 2302, container or pod 2404, insertinginto pod 2404, and can be provided to a method of transport 5524, in anexemplary embodiment. The centralized histology lab 5504 can includecombination 5510, including accessioning management software and/or userinterface, and transport/integrity verification processing, in anexample embodiment. The accessioning management software, in an exampleembodiment, can provide standard transport (TP) 5512, and embed, cut,stain, slide review and interpretation processing 5514, in an exampleembodiment. Combination 5510 can provide output to processing 5516,which can, for each cold box, provide processing including, e.g., butnot limited to, unloading pod, unloading container, inspecting for noleaks and/or no anomaly, unloading logger, reading/inspecting for enoughtime, and/or no anomaly, and if valid transport, accession sample andproceed to TP, or else reject the sample, or store in the cold box 2200for a longer time, in an example embodiment. From 5516, output can beprovided to 5518 including a method of data transport, such as, e.g.,but not limited to, USB-stick, tablet, uploading to vantage, etc., in anexample embodiment.

FIG. 25 depicts an exemplary cold tissue kit including, e.g., but notlimited to, an exemplary box 2220 kit 2200, shown with, the specimenvials 2402, specimen lids 2402, pod casings 2404 (discussed withreference to FIG. 4A), and exemplary TEMPOD datalogger 2406 (discussedwith reference to FIG. 4B) separate, without being inserted yet into thefoam inserts 2210, 2212 (shown, but not labeled, discussed further withreference to FIG. 3), along with cassettes 2304, 2306, according to anexemplary embodiment.

FIG. 26A and FIG. 26B are an example operating room system workflow 5300and an example AP Lab workflow 5320, respectively, according to anexemplary embodiment.

FIG. 26A illustrates an example system workflow 5300 for an operatingroom beginning with 5302. In 5302, formalin is stored in a refrigerator,in an exemplary embodiment. Formalin, a custom fixative is availablefrom Ventana, and may be prefilled in containers such as vials 2302 withlids 2402, and which may be stored in a cold temperature controlled areasuch as a refrigerator to be ready for use, in an exemplary embodiment.From 5302 workflow 5300 can continue with 5304.

In 5304, during surgery in the operating room, samples can be surgicallyremoved (e.g., 1-3 times/day/site), in an exemplary embodiment. Up tomultiple samples per patient (e.g., but not limited to, 1-2, or more)are possible, in an exemplary embodiment. A patient-identifier (ID) canbe read, e.g., but not limited to, from a patient's wristband, in anexemplary embodiment. From 5304, workflow 5300 can continue with 5306.

In 5306, tissue can be placed into the container or vial 2302, 2402, inan exemplary embodiment. The specimen can be inserted into a standardhistology cassette 2306, and the cassette 2306 can then be placed insideand held inside the vial 2302, 2402 container, in an exemplaryembodiment. Each container (vial 2302, with lid 2402) and cassette 2306can have a unique barcode identifier (ID), in an exemplary embodiment.From 5306, workflow 5300 can continue with 5308.

In 5308, the sample container (vial 2302 with lid 2402) can betransferred to a pod 2404, in an exemplary embodiment. The user of thecontainer can immediately transfer the vial 2302 with closed lid 2402 tothe pod 2404 as shown in FIG. 4A. The pod 2404 can also hold anexemplary datalogger 2406 in close proximity. The data logger 2406 canbe manually started to log time and temperature, in an exemplaryembodiment. In another exemplary embodiment, the data logger canautomatically start, e.g., but not limited to, at insertion into theslot in the pod, etc. Together, the pod 2404, vial 2302 (with lid 2404)and datalogger 2406 can be referred to as pod 2218. From 5308, workflow5300 can continue with 5310.

In 5310, the pod 2218 can be inserted into an exemplary cold box 2220for storage, as illustrated in diagram 2200 of FIG. 26, in an exemplaryembodiment. Storage in the box 2220 ensures that samples are kept at adesired temperature (e.g., but not limited to 4° C.) during transport,according to an exemplary embodiment. At the end of a surgical day, inone embodiment, the box of diagram 2200 may be shipped. From 5310,workflow 5300 can continue with 5312.

In 5312, the box 2220 with complete assembly can be transported to an APlab, and temperature can be recorded continuously during transport toensure requisite conditions of the sample are maintained, in anexemplary embodiment. From 5312, workflow 5300 can continue with 5314.

FIG. 26B illustrates an exemplary AP lab system workflow 5320, and maybegin with 5314 and may continue immediately with 5322, in an exemplaryembodiment.

In 5322, data may be read into the AP lab system, in an exemplaryembodiment. In an exemplary embodiment, upon arrival at the lab, a timeand temperature profile may be read from each pod's 2218 datalogger 2406an the integrity of the specimen can thereby be verified. Additionalcold storage times may be required for some samples, depending ondesired processing, in one embodiment. From 5322, workflow 5320 cancontinue with 5324.

In 5324, exemplary grossing can be performed, wherein the sample can bereviewed and then transferred, or kept, in a final cassette with barcodefor identification, in an exemplary embodiment. From 5324, workflow 5320can continue with 5326.

In 5326, exemplary processing of the sample can be performed, using anexemplary standard processor with an exemplary heated formalin step(e.g., but not limited to, 2 hours at 45° C.), tissue can besubsequently processed with typical alcohols/xylene/paraffin steps,according to an exemplary embodiment. From 5326, workflow 5320 cancontinue with 5328.

In 5328, exemplary stain and review can be performed, in an exemplaryembodiment. According to an exemplary embodiment, the processed samplescan be embedded, sectioned, and stained with various markers/probes. Thestain results can be reviewed and scored by pathologists, in anexemplary embodiment. Results for an example sample can then be reportedto a clinical database, according to an exemplary embodiment. From 5328,workflow 5320 can end, according to an exemplary embodiment.

FIGS. 27A and 27B illustrate two exemplary sealing options andvariations for an exemplary specimen cup.

FIG. 27A a wrap cup in parafilm 2800.

FIG. 27B illustrates in diagram 2810 a spill-protected sample insertionthrough a split septum, in the exemplary embodiment. In FIG. 28B, anexemplary fume reduction concept 2810 can include inserting an examplecassette 2304 coupled to a lid through an exemplary septum having one ormore slot(s) 2806, 2808 of a membrane seal to be placed in the fixativeof an example vial 2804, according to an exemplary embodiment. Accordingto exemplary embodiments of FIGS. 6A, 6B, an exemplary means to insertand retain an example standard histology cassette can be provided, suchthat the cassette upon insertion into a vial 2302 is submersed in fluidindependent of the orientation of the vial 2302 or pod 2404, accordingto an exemplary embodiment.

An exemplary clip feature 3504 (discussed further with reference toFIGS. 31A-31C, 32A-32B, 33A-33D, and 35A-35I) can be used to hold andretain cassettes 2304 with a corresponding feature 3504, according to anexemplary embodiment.

An exemplary carrier feature or cassette holder of FIGS. 33A-33D allowsa cassette 2304 to slide in and retain a cassette, according to anexemplary embodiment.

An exemplary embodiment can include an example dual sided lid, set forthin FIGS. 28, 29, 30A-30E, 31A-31C and 32A-32B, can hold an unusedcassette upward to avoid contamination with fixative prior to sampleinsertion, and easy handling, according to an exemplary embodiment. Theother side of the lid sealing the fluid inside the vial duringtransport/handling and to avoid spills/fumes, according to an exemplaryembodiment. After sample insertion into the cassette 2304 the lid can bereversed and screwed onto the vial with the fluid/fixative, according toan exemplary embodiment.

FIG. 28 is an illustration 2900 of an integrated cup design including anexemplary two sided lid 2902 shown with a cassette vertical orientation,in an exemplary embodiment. According to an exemplary embodiment, acarrier and a cup cap can be a single part (shown as different shades ofcolor), including a vial base portion, and a cap with cassette coupledto the cap, in an exemplary embodiment. According to an exemplaryembodiment, a single piece 4 well cup insert can be provided, howeverdifferent numbers of well cups may also be provided, in an exemplaryembodiment. An exemplary double cap concept is shown in“ship-to-hospital” position (i.e., with no tissue samples loaded in thecassettes yet), in an exemplary embodiment. An exemplary protectivesheath (not shown) around the cassette can be provided to protectagainst contamination and/or dust, in an exemplary embodiment.

FIG. 29 is an illustration 3000 of an integrated cup designillustrating, in the foreground, a reversed cap 3002 direction immersinga sample cassette 2306 in fixative (e.g., formalin in an exemplaryembodiment) in the vial 2302.

The exemplary unit can be shipped with fixative (e.g., formalin),already in cups with caps, in an exemplary embodiment. According to anexemplary embodiment, the technician can remove protective sheath (notshown), and can unscrew the two sided lid and can load the sample intocassette. In an example embodiment, the unit does not require thecassette to be removed from the cap. Once the cassette is loaded, thetechnician can reverse the cap direction and can screw onto the cupimmersing the sample in formalin, in an exemplary embodiment. Thetechnician can tell at a glance which cups have samples and which donot, in an exemplary embodiment. The cap, in an exemplary embodiment, istall enough (in its outer periphery) for label/barcode to be applied.The example sample cup cannot be left outside cold box, in an exemplaryembodiment.

FIG. 30A illustrates a method of use 3100 of the unit of FIGS. 29 and30, showing the last sample being loaded, according to an exemplaryembodiment.

FIG. 30B is an image 3110 illustrating an example cassette 2306, 2308with a slanted portion for labeling. In one example embodiment, thecassettes can be coupled to the cap in the vertically opposite way thanshown in FIG. 30A, for the barcode reader to more easily scan theslanted area, that is, with slanted side of the cassette facing downinto the vial during insertion, according to an exemplary embodiment.

FIG. 30C is an illustration 3120 of samples loaded and ready forshipment to a lab, according to an exemplary embodiment.

FIG. 30D is a partial cutaway view 3130 of the image of FIG. 30C,according to an exemplary embodiment.

FIG. 30E is a protected version 3140 of the vial container, according toan exemplary embodiment. A formalin filled cup can stay attached and canbe pre-filled (fits in the insert for shipment), according to anexemplary embodiment. A membrane that gets punctured when you screw itinto the carrier insert, gets the same type effect, according to anexemplary embodiment. The formalin from the now upside down cup can flowinto the lower insert cup, according to an exemplary embodiment. Thelower container can then protect the cassette from contamination,according to an exemplary embodiment.

FIG. 31A illustrates individual components of an exemplary cassetteholder 3200 (with top view 3202, and bottom perspective view 3204showing a cassette clip/holder 3204, an exemplary o-ring 3208 (e.g.,solvent compatible), and one or a plurality (e.g., four) snap clips(e.g., male) 3206.

FIG. 31B illustrates an exemplary cassette receiver 3210, with a vialscrew top including in a top view a pierceable septum 3212, one or aplurality (e.g., 4) of snap clip receivers (female), and in a bottomview a threading (e.g., female) 3218 for mating with the vial, and ano-ring (e.g., solvent compatible).

FIG. 31C illustrates a fixative vial 3220, with threaded vial top (e.g.,male) 3222, a void volume 3224 of approximately the volume of thecassette, and a fixative (e.g., a formalin solution) 3226. The vial 3220and cassette receiver 3210 can be combined prior to shipping, asillustrated.

FIG. 32A illustrates a user receiving 3220 the cassette holder 3200.

FIG. 32B illustrates in diagram 3240 the cassette receiver/vial assembly3240 of cassette receiver 3210, and vial 3220. The septum lid can becovered during shipment to protect from undesired piercing, according toan exemplary embodiment. In one embodiment, the cover can be adisposable clip-on lid or protected shipping conditions.

FIG. 33A illustrates an exemplary process 3300 of attaching the cassette3302 to the cassette holder 3200 at holder 3204, forming a cassetteassembly.

FIG. 33B illustrates an exemplary process 3310 of piercing with thecassette assembly of FIG. 33A, the septum of the cassette receiver3210/vial 3220 assembly 3240.

FIG. 33C illustrates an exemplary process 3320 of completely sealing avial to be ready for transport.

FIG. 33D illustrates an exemplary process 3330 of having the receivinglab unscrew the cassette holder/receiver assembly and removing thecassette assembly from the vial, as shown. The process of the series ofprocess steps of FIG. 33A-33D include that most parts can be plasticmolded, that there is reduced exposure to formalin vapors, that thecassette remains completely submerged during transport, that thecontainer can ship ready to use, and that the process is compatible withgeneric cassettes.

FIG. 34A illustrates a first state with a sample in the air, exposed tothe environment, with a wedge arm 3402, lid 3200, and cassette 3302,with protective cylinder 3404, where the wedge arm 3402 can break amembrane when the sample is placed into the vial 3220.

FIG. 34B illustrates in diagram 3410 a fixative vial 3220 with abreakable membrane 3412.

FIGS. 34C and 34D illustrate in 3420 and 3430, respectively, acontinuation of the first state of FIGS. 34A and 34B with the samplecontainer lid 3200 of FIG. 34A affixed to the protective cylinder 3404,and with the fixative container vial 3220 of FIG. 34D still isolated andwith membrane 3412 still intact. The inner diameter of 3404, is slightlygreater than the outer diameter of vial 3220, to permit placing one atopthe other as shown in FIG. 34E.

FIG. 34E illustrates in diagram 3440 a transition to a second state. Byscrewing the sample container 3200, 3404 over and around the fixativecontainer 3220, isolating the sample from the environment, the fixativewill still remain isolated prior to the arm breaking the membrane 3412by the arm 3402 when inserting the cassette 3302 into the vial 3220.

FIGS. 35A-35I illustrate an exemplary fume reduction feature explained,with an exemplary cassette cover in an exemplary dry implementation.

FIG. 35A includes a diagram 3500 including a tissue container vial 3520with a cassette septum 3510 membrane for isolating the fixative, with anexample split septum 3514, a tissue container lid 3502 with an exampleconcave portion 3506 for receiving an example cassette clip 3504, whichcouples a tissue cassette 3302 to the lid 3502, and a cassette cover3508 which can isolate the tissue cassette 3302 by covering the cassetteand coupling with an edge 3510 about a rim of tissue container lid 3502,in an example embodiment.

FIG. 35B depicts a diagram 3510 illustrates a cassette 3302 is dry andsterile above the lid 3502 and the container 3520, which can bepre-filled with fixative, such as, e.g., formalin, in an exampleembodiment.

FIG. 35C depicts a diagram 3520 illustrates a cassette cover can beremoved and discarded breaking a sterile seal between the cover 3508 andlid 3502 at edge 3510 of the lid.

FIG. 35D depicts a diagram 3530 a cassette 3302 and clip 3504 unscrewedfrom the lid, together, in an example embodiment. In an exampleembodiment, the clip 3504 may be threaded and mate with inner threads ofconcavity 3506 of lid 3502. In another embodiment, the clip can bescrewed to the lid 3502, or snapped into the lid 3502, in an exampleembodiment.

FIG. 35E depicts a diagram 3540 illustrating tissue being inserted intoan example cassette 3302 coupled to cassette clip 3504, in an exemplaryembodiment.

FIG. 35F depicts a diagram 3550 illustrating removing tissue containerlid 3502 from tissue container vial 3520, and then screwing/coupling thecassette clip with tissue cassette into the inside of an example doublesided lid with threads inside, as well as outside of lid 3502, in anexemplary embodiment.

FIG. 35G depicts a diagram 3560 illustrating inserting the tissuecassette 3302, clip 3504 and lid 3502 cassette assembly into thecontainer 3520 through cassette septum 3512 into the container 3520, inan exemplary embodiment.

FIG. 35H depicts a diagram 3570 illustrating the tissue and cassetteplaced sealed in the tissue container vial 3520 filled with fixative(e.g., formalin), in an exemplary embodiment.

FIG. 35I depicts a diagram 3580 illustrating the septum 3512 can beremovable with the cassette 3302, clip 3504, and lid 3502 to provide asplash-free, easy to access to the cassette 3302 system, in an exemplaryembodiment.

FIGS. 36A-36I (collectively, FIG. 36) is an exemplary preanalyticalworkflow beginning with FIG. 36A diagram 3600.

FIG. 36A depicts a diagram 3600 illustrating a tissue container withformalin fixative stored at 4° C., in an exemplary embodiment.

FIG. 36B depicts a diagram 3610 illustrating tissue collection, in anexemplary embodiment.

FIG. 36C depicts a diagram 3620 illustrating a start or initiation ofdata collection, in an exemplary embodiment.

FIG. 36D depicts a diagram 3630 illustrating pairing tissue with datacollection, in an exemplary embodiment.

FIG. 36E depicts a diagram 3640 illustrating preparing multiple samplesfor shipping, in this case an example four pods at a time, in anexemplary embodiment.

FIG. 36F depicts a diagram 3650 illustrating packaging in example coldbox transport box 2200 (also shown in FIG. 26), in an exemplaryembodiment.

FIG. 36G depicts a diagram 3660 illustrating shipping to centralfacility for processing, in an exemplary embodiment.

FIG. 36H depicts a diagram 3670 illustrating data collection hand-off ofthe datalogger and sample cassette for central processing, in anexemplary embodiment.

FIG. 36I depicts a diagram 3680 illustrating an example fixationprocessor collects transport data, performs any desired final fixationsteps included in a fixation protocol, and provides a comprehensivefixation quality report for each tissue sample, in an exemplaryembodiment.

FIGS. 37-44, 7 and 8 illustrate example specimen vials, lids, clips, andcassettes, of various example embodiments.

FIG. 37 is an exemplary exploded view 3700 of an exemplary tissue potwith lid, and exemplary 10% NBF. The container vial 3520 can includefixative 3712 therein. As shown, formalin 10% NBF can be pre-filled andcooled to a desired temperature. The cassette 3302 can be coupled toclip 3504. Clip 3504 can be screwed in, as shown in diagram 3700 and cancouple the clip 3504 to cylinder 3708, which can be screwed to lid 3200with o-ring 3706 and cap 3702 with screws 3704 through lid 3200.

FIG. 38 is an exemplary exploded view of an exemplary tissue pot withlid, and exemplary 30% NBF. The container vial 3520 can include fixative3812 therein. As shown, formalin 10% NBF can be pre-filled and cooled toa desired temperature. The cassette 3302 can be coupled to clip 3504.Clip 3504 can be screwed in, as shown in diagram 3800 and can couple theclip 3504 to cylinder 3808, which can be screwed to lid 3200 with o-ring3806 and cap 3802 with screws 3804 through lid 3200.

FIG. 39A is a side view 3900 and FIG. 39B is a top view 3910 of anexemplary CELLSTOR lid 3200 modification, of an exemplary specimen lid3200 with exemplary three screw holes there through, in an exampleembodiment.

FIGS. 40A-40D are various views of an exemplary cell block cassette clip3504, having an exemplary angled slot for clipping to an exemplarycassette. FIG. 40A is a front side orthographic view 4000, FIG. 40B isan example top orthographic view 4010 of the clip 3504. FIG. 40C is anexample right edge side orthographic view 4020. FIG. 40D is an exampleisometric perspective view 4030 of clip 3504.

FIGS. 41A-41D, and 42A-42B are various views of an exemplary cell blockcassette. FIG. 41A is a bottom orthographic view 4000 of cassette 3302.FIG. 41B is a side edge orthographic view 4010 of cassette 3302. FIG.41C is a top orthographic view 4020 of cassette 3302. FIG. 41D is aright side edge orthographic view 4030 of cassette 3302.

FIG. 42A is a top isometric perspective view 4200 of cassette 3302.

FIG. 42B is a bottom isometric perspective view 4210 of cassette 3302.

FIGS. 43A-43C are various views of an exemplary cell transport cap plate3702, 3802, configured as a cap for the specimen lid 3200. FIG. 43A is atop orthographic view 4300 of cap plate 3702, 3802. FIG. 43B is a topedge orthographic view 4310 of cap plate 3702, 3802. FIG. 43C is a frontisometric perspective view 4320 of cap plate 4320.

FIGS. 44A-44E are various views of an exemplary cell transport sealblock 3708, 3808, which resembles a straight screw head, including anexemplary slotted cassette clip of one embodiment, as can be used in theprocess set forth in FIGS. 35A-35I.

FIG. 44A is a front orthographic view 4400 of cell transport seal block3708, 3808. FIG. 44B is a top orthographic view 4410 of cell transportseal block 3708, 3808, with partial cutaway cross-section A-A. FIG. 44Cis a right side orthographic view 4420 of cell transport seal block3708, 3808. FIG. 44E is a back orthographic view 4440 of cell transportseal block 3708, 3808. FIG. 44D is an isometric perspective view 4430 ofexemplary cell transport seal block 3708, 3808.

Another exemplary embodiment can include a single sided version of thelid, where the cassette is held separately until the sample is ready forinsertion, according to an exemplary embodiment.

An exemplary embodiment can include data logging electronics 2406, 2408,which can be held separately from the vial 2302 via a POD 2404construct, with thermal communication to allow monitoring by the datalogger 2406, 2408 of the vial and its sample contents.

An exemplary embodiment can provide an exemplary timer/clock functionassociated with the insertion of the container, and can log other timesand/or temperatures of events, or at specific and/or periodic times.

An exemplary embodiment can provide an exemplary feature having a meansto indicate completion of a predetermined time after the insertion ormanual start were established.

An exemplary embodiment can provide an exemplary display, monitor orindicator (such as, e.g., with an indicating/monitoring/displayingand/or storing system and/or method LED/LCD).

An exemplary embodiment can provide an exemplary feature having atemperature recording capability.

An exemplary embodiment can provide an exemplary feature of having ameans to record and indicate temperature excursions againstpreprogrammed thresholds (heating, freezing), indicating with LED/LCD.

An exemplary embodiment can provide an exemplary feature having a meansto monitor temperature excursions and time after sealing of thecontainer, and can indicate successful completion per LED/LCD againstpreprogrammed thresholds.

An exemplary embodiment can provide an exemplary feature including oneor more memories to store and/or include details about the sample ID,location relative to WiFi transponders, and/or record date/time, etc.

An exemplary embodiment can provide an exemplary means to communicatevia RFID/Bluetooth/NFC/Wireless/Zig Bee or direct connection to aPC/tablet/Cell Phone.

Finally, for an overview of the pre-analytical workflow, the reader isdirected further to the detailed discussion with reference to FIGS.36A-36I, above, according to one example embodiment.

In one embodiment, the transport systems demonstrated herein are usefulin an “autonomous heating” embodiment. As used herein, an autonomousheating embodiment is an embodiment in which the transport device isconfigured to hold the fixative solution within a first cold temperaturerange (typically in the range of 0° C. to 7° C.) for a period of timesufficient to allow the fixative to diffuse throughout the tissue sample(typically at least 2 hours). After diffusion has completed, thetemperature of the transport device is gradually allowed to rise toambient temperature. As the temperature rises, the fixation reactionbegins to proceed, thereby inducing fixation of the tissue sample. Anexemplary assembly is illustrated at FIG. 45A-45C. An insulated box(4501) is provided, having a cavity (4502) sized to accept a data logger(4503), such as a TEMPOD data logger (TempSen). Preferably, the datalogger is pre-chilled to a temperature in the range of 0° C. to 7° C.The data logger (4503) is configured with a cavity (4504) sized toaccept a cooling block (4505) containing a phase change medium having amelting temperature of ˜4° C. (although in principle, any meltingtemperature in the range of 0° C. to 7° C. could be used). The coolingblock includes a cavity (4506) sized to accept a sample container (4507)holding a tissue sample and a volume of a fixative solution (preferablypre-chilled in a range of 0° C. to 7° C.). To demonstrate theeffectiveness of such a device, Calu3 xenographs were stored in 10%neutral buffered formalin in a transport device as described herein at15° C., 19° C., 21° C., and 23° C. over a time course. The temperatureof the NBF is shown at FIG. 46A. The resulting fixed tissue samples wereimmunhistochemically stained for pAkt. Results are shown at FIGS. 46Band 46C. A 2+2 control (2 hours at 4° C. in NBF, followed by 45° C. inNBF) is provided as a positive staining sample. As can be seen, theexemplary device preserves pAkt to a comparable extent as the 2+2protocol.

In one simple embodiment, an autonomous heating embodiment is providedin which time the data logger is replaced by a timer apparatus whichuses diffusion of a colored liquid through an absorbent membrane as atimer to indicate how long the colored liquid has been in contact withthe absorbent membrane. Measurements of the time of arrival of thecolored liquid front at various points along the length of the absorbentmembrane can be used as a rough measurement of the time that a tissuesample has been in fixative. In some embodiments, the timer apparatus iscapable of determining the amount of time that the fixative has spent ata particular temperature range. For example, a device for monitoring thetime that a tissue sample has spent in room temperature fixative cancomprise an absorbent membrane separated from a dye-soaked pad by adye-impervious film. Once the film is removed, the dye comes intocontact with the absorbent membrane and starts to diffuse along itslength. The distance that the dye is seen to have diffused down themembrane indicates the length of time since the film was removed. Textand markings on the front of the device can either show the user howlong the tissue has been in fixative, or give an indication of the stateof the tissue. A similar device can measure the time that a tissuesample has spent at cold (e.g. 0-7° C.) and room temperature. A firstcolored dye indicates the length of time spent at cold temperature bythe same method of diffusion along an absorbent membrane and isinitiated by removal of the dye-impervious barrier. When the temperatureincreases beyond the desired temperature range, the dye should stopdiffusing and thus give a record of how long the tissue sample was at0-7° C. A second colored dye, now mixed with a material which melts at aspecific temperature above the cold temperature range (e.g. roomtemperature), will only diffuse along a separate absorbent membrane whenthe environment around the tissue sample reaches the specifictemperature. Thus one can also monitor the time that the tissue samplewas at or above the specific temperature. Since two times are beingmonitored, there are several possible states for the device to end in,indicating whether the tissue sample was exposed to both coldtemperature and a higher temperature for the proper amount of time forideal fixation. It is also possible for the device to merely report thelength of time (in hours) at each temperature. The device for monitoringthe time spent in cold and room temperature includes a method to haltthe diffusion of the first dye through the absorbent membrane when thetemperature increases in order to provide a record of how long thetissue sample spent in the cold temperature. For example, a phase changematerial is introduced behind the absorbent membrane in contact with thefirst dye. This phase change material (PCM) will remain solid at coldtemperature, but when the ambient temperature increases it will melt.The chemical composition of the PCM, absorbent membrane and first dyeshould be chosen such that the PCM fouls the absorbent membrane andprevents further diffusion of the first dye. For instance, the PCM couldbe a paraffin which melts at 18° C., and the first dye could be amixture of dye, water and glycerol. In that case the hydrophobicity ofthe paraffin absorbed into the membrane would prevent the aqueousmixture from diffusing further along the absorbent membrane. Exemplaryarrangements are illustrated at FIGS. 45A-45D. An advantage of such anembodiment is that the entire apparatus may be presented in aninexpensive, disposable format.

The above detailed descriptions of embodiments of the technology are notintended to be exhaustive or to limit the technology to the precise formdisclosed above. Although specific embodiments of, and examples for, thetechnology are described above for illustrative purposes, variousequivalent modifications are possible within the scope of thetechnology, as those skilled in the relevant art will recognize. Forexample, while steps are presented in a given order, alternativeembodiments may perform steps in a different order. The variousembodiments described herein may also be combined to provide furtherembodiments. The analyzers, processing systems, controllers, andcomputing devices disclosed herein can include memory for storinginformation. Such memory can be computer-readable media that may beencoded with computer-executable instructions that implement thetechnology, e.g., a computer-readable medium that contains theinstructions. In addition, the instructions, data structures, andmessage structures may be transmitted via a data transmission medium,such as a signal on a communications link and may be encrypted.Accordingly, computer-readable media include computer-readable storagemedia upon which data can be stored and computer-readable transmissionmedia upon which data can be transmitted. The data can include, withoutlimitation, object classifier routines, ground truth slide data (orother types of reference images), reference images, segmentationroutines, scoring protocols, or the like. Various communications linksmay be used, such as the Internet, a local area network, a wide areanetwork, a point-to-point dial-up connection, a cell phone network, andso on. It should be noted that, as used in this specification and theappended claims, the singular forms “a,” “an,” and “the” include pluralreferents unless the content clearly dictates otherwise. Thus, forexample, reference to “a tissue sample” includes a single tissue sample,or two or more tissue samples. It should also be noted that the term“or” is generally employed in its sense including “and/or” unless thecontent clearly dictates otherwise. Also, as used herein, the term“comprises” is used in its legally accepted definition and manner, andnothing herein is intended to change such meaning. The systems andmethods disclosed herein can be used with tissue samples from mammals.As used herein, the term mammal includes both human and non-humanmammals. Similarly, the term “subject” includes both human andveterinary subjects.

FURTHER EMBODIMENTS

1. A portable transporter system for protecting biomarkers in a tissuesample, comprising:

-   -   a specimen container configured to retain or hold at least one        tissue sample in a reagent, wherein the reagent is chilled to        from about 0° C. to about 15° C. prior to transportation to a        laboratory;    -   a carrier assembly configured to retain or hold the specimen        container; and    -   a monitoring system that comprises at least one sensor        configured to obtain, store or transmit one of or both time and        temperature information about the tissue sample and/or reagent        in the specimen container when the specimen container is        positioned in the carrier assembly and transported to a        laboratory without directly contacting the contents of the        specimen container, wherein the monitoring system further        comprises a data logging device.

2. The transporter system of embodiment 1 wherein the reagent is atissue fixation reagent.

3. The transporter system of embodiment 1 further comprising a tissuesample cassette for retaining one or more tissue samples therein andwherein the specimen container is configured to retain and hold thetissue sample cassette in the reagent.

4. The transporter system of embodiment 1, wherein the monitoring systemmeasures the temperature of the reagent and/or tissue sample in thespecimen container over a period of time.

5. The transporter system of embodiment 1, further including aninsulator and/or a cooling element capable of maintaining the volume ofthe formalin-based fixative solution at a temperature in the range of 0°C. to 15° C. for at least 2 hours under shipping conditions when thetissue sample transport and storage assembly is assembled.

6. The transporter system of embodiment 1, wherein the monitoring systemis physically coupled to the carrier assembly.

7. The transporter system of embodiment 1, wherein the monitoring systemis electronically coupled to the carrier assembly.

8. The transporter system of embodiment 1, wherein the monitoring systemcomprises at least one sensor, wherein the sensor is a temperaturesensor positioned in the specimen container and comprises a transmitterthat outputs a temperature signal to a receiver located in the carrierassembly holding that specimen container.

9. The transporter system of embodiment 1, wherein the monitoring systemeither stores temperature measurements indicative of the temperature ofthe reagent and/or the tissue sample or transmits such information to anexternal receiver wherein the receiver comprises a memory device.

10. The transporter system of embodiment 4, wherein the specimencontainer comprises a temperature sensor wherein said sensor transmitsthe temperature of the reagent contained therein to the monitoringsystem.

11. The transport system of embodiment 1, wherein the specimen containerfurther comprises a radio frequency identification (RFID) tag and themonitoring system further comprises a RFID reader.

12. The transport system of embodiment 1, further comprising a transportcontainer for transporting the specimen container, carrier assembly andmonitoring system wherein the transport container comprises an ice bath.

13. The transport system of embodiment 1, further comprising a transportcontainer for transporting the specimen container, carrier assembly andmonitoring system, wherein the transport container is configured tomaintain the temperature of a specimen container therein to from about0° C. to about 15° C. for up to 14 days.

14. The transporter system of embodiment 1, further comprising aplurality of carrier assemblies and a plurality of specimen containerswherein each of the plurality of carrier assemblies holds a separate oneof the plurality of specimen containers.

15. The transporter system of embodiment 1 further comprises a coolingdevice configured to reduce or maintain an average temperature ofreagent in the specimen container to or at a temperature equal to orlower than about 15° C.

16. An apparatus for transporting tissue samples, comprising:

-   -   a plurality of specimen containers holding reagents;    -   a plurality of carrier assemblies, each assembly comprising a        specimen container holder and a sensor assembly positioned to        monitor the temperature of the specimen container or the reagent        held in each specimen container and time the container is held        in the holder, wherein the sensor assembly comprises a        temperature sensor and a time sensor;    -   a transport assembly for carrying the multiple carrier        assemblies; and    -   a transporter for carrying at least one transport assembly.

17. The apparatus of embodiment 16, wherein the sensor assembly of eachof the plurality of carrier assemblies comprises a temperature sensorpositioned within the specimen container, wherein the temperature sensorincludes a transmitter that outputs a temperature signal to a receiverlocated in the carrier assembly holding that specimen container.

18. The apparatus of embodiment 17, further comprising a computingdevice communicatively coupled to ultrasound transmitters and ultrasoundreceivers, wherein the computing device is configured to evaluate soundspeeds in the tissue sample by evaluating time of flight changes ofacoustic waves from one or more of the transmitters.

19. The apparatus of embodiment 18, wherein the computing device isfurther configured to compensate for changes in the time of flight ofacoustic waves in a fixative or sample associated with environmentalchanges.

20. The apparatus of embodiment 17, further comprising the computingdevice containing instructions, when executed, to cause the computingdevice to compensate for temperature changes of one or more of thereagent or sample.

21. A transporter system for carrying a tissue sample, comprising:

-   -   a specimen container configured to hold a tissue sample in a        fixative;    -   a carrier assembly configured to retain or hold a specimen        container;    -   a monitoring system that obtains and stores time and temperature        information about the tissue sample in the specimen container        when the specimen container is positioned in the carrier        assembly; and    -   a transport container configured to hold the carrier assembly        holding the specimen container and monitoring system, wherein        the transport container is further configured to maintain the        temperature of the internal holding chamber at from about 0° C.        to about 20° C. for at least 1 hour.

22. The transporter system of embodiment 21, wherein the transportcontainer is configured to maintain the temperature of the internalholding chamber at from about 0° C. to about 15° C. for at least 24hours.

23. The transporter system of embodiment 21, wherein the transportcontainer is configured to maintain the temperature of the internalholding chamber at from about 0° C. to about 15° C. for up to 14 days.

24. The transporter system of embodiment 21, wherein the carrierassembly is configured to hold a plurality of specimen containers eachholding a respective tissue sample, wherein the monitoring systemincludes a plurality of temperature sensors each measuring a temperatureof the tissue sample in one of the specimen containers.

25. The transporter system of embodiment 24, wherein the monitoringsystem measures the temperature of the fixative and/or tissue sampleover a period of time.

26. The transporter system of embodiment 21, wherein the monitoringsystem includes a temperature sensor carried by the specimen containerand positioned to measure a temperature of the fixative.

27. The transporter system of embodiment 21, wherein the monitoringsystem stores temperature measurements indicative of the temperature ofthe fixative and/or the tissue sample.

28. The transport system of embodiment 21, wherein the monitoring systemincludes a reader physically coupled to the carrier assembly and aradio-frequency identification tag coupled to the specimen container.

29. The transporter system of embodiment 21, wherein the transportcontainer comprises a cooling device configured to reduce an averagetemperature of fixative in the specimen container from room temperatureto a temperature equal to or lower than about 15° C.

30. The transporter system of embodiment 21, wherein the monitoringsystem further comprises a timer.

31. The transporter system of embodiment 21, wherein the specimencontainer includes a cover and a main body, wherein the cover includes aholder configured to hold the tissue sample in contact with thefixative.

32. A method for processing a tissue sample, comprising:

-   -   contacting the tissue sample with a fixative chilled to from        about 0° C. to about 15° C., wherein the fixative is contained        in a specimen receiving container;    -   placing a specimen container in a holding well of a carrier        assembly further comprising at least one data logging device;    -   detecting and storing time and temperature information        associated with the tissue sample in the data logger of the        carrier assembly;    -   placing the carrier assembly in a transport assembly; and    -   transporting the transport assembly to a laboratory.

33. The method of embodiment 32, wherein detecting the time andtemperature information associated with the tissue sample includes:

-   -   measuring a temperature of the fixative and/or the tissue        sample, and    -   measuring a contact period of time in which the fixative        contacts the tissue sample.

34. The method of embodiment 32, wherein detecting the time andtemperature information associated with the tissue sample includesobtaining temperature measurements and storing the temperaturemeasurements.

35. A self-cooling portable transporter system for carrying a tissuesample contacting a fixative, comprising:

-   -   a transport container including a holding chamber;    -   a fixative in the holding chamber; and    -   a fixation-inhibiting cooling device carried by the transport        container and in thermal communication with the fixative in the        holding chamber, wherein the fixation-inhibiting cooling device        is operable to reduce a temperature of the fixative to inhibit        fixation of a tissue sample contacting the fixative.

36. The transporter system of embodiment 35, further comprising amonitoring system carried by the transport container and configured toobtain and store time-temperature information about the tissue samplethrough which the fixative diffuses after the temperature of thefixative has been reduced by the fixation-inhibiting cooling device.

37. The transporter system of embodiment 35, further comprising amonitoring system that obtains temperature over time information aboutthe tissue sample, the fixative, or both.

38. The transporter system of embodiment 35, wherein thefixation-inhibiting cooling device is configured to reduce thetemperature of about 100 ml to 400 ml of fixative by at least about 25°C.

39. The transporter system of embodiment 35, wherein the transportcontainer has an open configuration for receiving the tissue sample anda closed configuration for sealing the holding chamber, wherein thefixation-inhibiting cooling device has a non-cooling mode and a coolingmode, and wherein the fixation-inhibiting cooling device in the coolingmode reduces a temperature of the fixative at least about 25° C. in lessthan about 5 minutes.

40. The transporter system of embodiment 39, wherein the transportcontainer is configured to maintain the temperature of the fixative ator below about 10° C. for at least one hour while an ambient temperaturesurrounding the transport container is equal to or higher than about 10°C.

41. The transporter system of embodiment 39, wherein the fixativecomprises formalin.

42. The transporter system of embodiment 35, wherein thefixation-inhibiting cooling device includes a first substance and asecond substance that mix to produce an endothermic reaction in responseto the fixation-inhibiting cooling device changing from a non-coolingmode to a cooling mode.

43. The transporter system of embodiment 35, wherein thefixation-inhibiting cooling device comprises a desiccant and a substancethat evaporates to cool the fixative.

44. The transporter system of embodiment 35, wherein thefixation-inhibiting cooling device includes a refrigeration unit or athermoelectric cooler unit.

45. The transporter system of embodiment 35, wherein thefixation-inhibiting cooling device is configured to cool the tissuesample to a temperature equal to or lower than about 4° C.

46. The transporter system of embodiment 35, wherein thefixation-inhibiting cooling device is configured to reduce a temperatureof about 100 ml of the fixative at least about 15° C. in less than about4 minutes.

47. The transporter system of embodiment 35, further comprising aspecimen container positioned in the holding chamber and holding thefixative.

48. A transporter system for carrying a tissue sample, comprising:

-   -   a transport container;    -   a specimen container configured to hold a tissue sample in a        fixative; and    -   a monitoring system that obtains and stores time-temperature        information about the tissue sample in the specimen container        when the specimen container is positioned in the transport        container.

49. The transporter system of embodiment 48, wherein the transportcontainer is configured to hold a plurality of specimen containers eachholding a respective tissue sample, wherein the monitoring systemincludes a plurality of temperature sensors each measuring a temperatureof the tissue sample in one of the specimen containers.

50. The transporter system of embodiment 48, wherein the monitoringsystem measures the temperature of the fixative and/or tissue sampleover a period of time.

51. The transporter system of embodiment 48, wherein the monitoringsystem includes a temperature sensor carried by the specimen containerand positioned to measure a temperature of the fixative.

52. The transporter system of embodiment 48, wherein the monitoringsystem stores temperature measurements indicative of the temperature ofthe fixative and/or the tissue sample.

53. The transporter system of embodiment 48, wherein the monitoringsystem includes a reader physically coupled to the transport containerand a radio-frequency identification tag coupled to the specimencontainer.

54. The transporter system of embodiment 48, wherein the transportcontainer contains a substance that vaporizes to reduce the temperatureof the fixative and/or the tissue sample.

55. The transporter system of embodiment 48, further comprising acooling device configured to reduce an average temperature of 100 ml offixative in the specimen container from room temperature to atemperature equal to or lower than about 4° C.

56. The transporter system of embodiment 48, further comprising acooling device configured to reduce an average temperature of 100 ml offixative in the specimen container at least 15° C. in less than about 4minutes.

57. The transporter system of embodiment 48, further comprising acooling device configured to reduce an average temperature of at leastone 1 liter of fixative in the specimen container by at least 15° C.

58. The transporter system of embodiment 48, wherein the monitoringsystem further comprises a timer.

59. The transporter system of embodiment 48, wherein the specimencontainer includes a cover and a main body, wherein the cover includes aholder configured to hold the tissue sample in contact with thefixative.

60. An apparatus for processing a tissue sample, comprising:

-   -   a plurality of processing containers holding reagents;    -   a carrier assembly including a specimen holder and a sensor        assembly positioned to acoustically analyze a tissue sample held        by the specimen holder when the tissue sample is submerged in        one of the reagents; and    -   a transport mechanism carrying the carrier assembly and operable        to submerge the sensor assembly and tissue sample held by the        specimen holder in the reagents in the containers.

61. The apparatus of embodiment 60, further comprising a computingdevice with instructions, when executed, that cause the apparatus to:

-   -   if diffusion of fixative through the tissue sample is at or        above a target diffusion level, a fixation process is performed        on the tissue sample; and    -   if diffusion of fixative through the tissue sample is below the        target diffusion level, a diffusion process is performed on the        tissue sample.

62. The apparatus of embodiment 60, wherein the sensor assembly includesa plurality of sensors positioned to acoustically analyze the tissuesample located in a detection zone, wherein each sensor includes atransmitter that outputs acoustic waves and a receiver that detects theacoustic waves from a respective one of the transmitters, and whereinthe detection zone is between the transmitters and the receivers.

63. The apparatus of embodiment 60, wherein the sensor assemblyincludes:

-   -   a plurality of transmitters, each of the transmitters outputs        acoustic waves; and    -   a plurality of receivers, each of the receivers is positioned to        detect the acoustic waves from a respective one of the        transmitters while the sensor assembly moves relative to the        sample to scan the sample in one direction.

64. The apparatus of embodiment 60, further comprising a computingdevice communicatively coupled to the transmitters and receivers,wherein the computing device is configured to evaluate sound speeds inthe tissue sample by evaluating time of flight changes of acoustic wavesfrom one or more of the transmitters.

65. The apparatus of embodiment 60, further comprising a computingdevice configured to compensate for changes with a fixative associatedwith environmental changes.

66. The apparatus of embodiment 60, further comprising the computingdevice containing instructions, when executed, cause the computingdevice to compensate for temperature changes of one or more of thereagents.

67. A method for processing a tissue sample, comprising:

-   -   contacting the tissue sample with a fixative;    -   moving a transport container containing the fixative and the        tissue sample from a first location to a second location while        an average temperature of the fixative is equal to or less than        about 5° C.; and    -   detecting time-temperature information associated with the        tissue sample while moving the transport container from the        first location and the second location.

68. The method of embodiment 67, wherein detecting the time-temperatureinformation associated with the tissue sample includes:

-   -   measuring a temperature of the fixative and/or the tissue        sample, and    -   measuring a contact period of time in which the fixative        contacts the tissue sample.

69. The method of embodiment 67, wherein detecting the time-temperatureinformation associated with the tissue sample includes obtainingtemperature measurements and storing the temperature measurements.

70. The method of embodiment 67, further comprising after contacting thetissue sample with the fixative, which is at an average temperatureequal to or lower than about 10° C. for at least about one hour, heatingthe tissue sample to a temperature from about 35° C. to about 45° C.

71. The method of embodiment 67, further comprising removing the tissuesample from the transport container; and performing a histologicalprocess on the tissue sample based on the time-temperature information.

72. A method for processing a tissue sample contacting a fixative heldin a specimen container, comprising:

-   -   removing a tissue sample from a specimen container;    -   transmitting acoustic waves through the tissue sample;    -   evaluating diffusion of fixative through the tissue sample based        on time of flight detection using acoustic waves; and    -   performing a histological process on the tissue sample based on        the evaluation of the diffusion of the fixative.

73. The method of embodiment 72, wherein evaluating the diffusion of thefixative includes monitoring time of flight of the acoustic waves thattravel through the tissue sample while the tissue sample contacts thefixative.

74. The method of embodiment 72, wherein evaluating the diffusion of thefixative includes monitoring time of flight of the acoustic waves thattravel through the tissue sample while fixative displaces interstitialliquid from the tissue sample.

75. The method of embodiment 72, further comprising contacting thetissue sample with the fixative (a) after removing the tissue samplefrom the specimen container and (b) while evaluating diffusion of thefixative in the tissue sample.

76. A portable transporter system adapted to carry a tissue sample in astandard histology cassette, the tissue sample contacting a fixative,comprising:

-   -   a transport container comprising:    -   a holding chamber;    -   a cassette septum; and    -   a fixative in said holding chamber; and    -   a container lid comprising a cassette clip attachably couplable        to receive, hold and retain the standard histology cassette.

77. The portable transporter system according to embodiment 76, furthercomprising: a fixation-inhibiting cooling device carried by thetransport container and in thermal communication with said fixative insaid holding chamber, wherein said fixation-inhibiting cooling device isoperable to reduce a temperature of said fixative to inhibit fixation ofthe tissue sample contacting said fixative.

78. The portable transporter system according to embodiment 76, furthercomprising a cassette cover removably coupled to said container lid.

79. The portable transporter system according to embodiment 76, whereinsaid cassette cover comprises a sterile seal between said cassette coverand said container lid.

80. The portable transporter system according to embodiment 76, whereinsaid fixative comprises formalin.

81. The portable transporter system according to embodiment 76, whereinsaid cassette septum is removable with said container lid, said cassetteclip, and the standard histology cassette.

82. The portable transporter system according to embodiment 81, whereinsaid removable cassette septum provides easy access to the standardhistology cassette while eliminating splashing of said fixative.

83. The portable transporter system according to embodiment 76, whereinsaid cassette clip is removable couplable to said container lid.

84. The portable transporter system according to embodiment 76, whereinsaid container clip is operable to be screwed into at least one side ofsaid container lid.

85. The portable transporter system according to embodiment 76, whereinthe cassette clip is operable to be clipped to the standard histologycassette.

86. The portable transporter system according to embodiment 76, whereinsaid cassette clip comprises:

-   -   a first end operable to be coupled to at least one side of said        container lid; and    -   a second end operable to be coupled to the standard histology        cassette.

87. The portable transporter system according to embodiment 76, whereinsaid cassette septum comprises a breakable membrane.

88. The portable transporter system according to embodiment 87, whereinsaid container lid further comprises an arm operable to break saidbreakable membrane upon insertion of the cassette into said holdingchamber of said transport container.

89. The portable transporter system according to embodiment 88, whereinsaid container lid further comprises a cylindrical portion comprising aninternal screw thread portion adapted to couple with an external screwthread portion of said transport container, operable to isolate thesample, air and said fixative from a surrounding environment.

90. The portable transporter system according to embodiment 76, furthercomprising a data collection device.

91. The portable transporter system according to embodiment 90, furthercomprising a coupler for coupling said data collection device to saidtransport container together to form a paired tissue sample with datacollection.

92. The portable transporter system according to embodiment 91, furthercomprising a first shipping receptacle portion for receiving a pluralityof said paired tissue samples with data collections.

93. The portable transporter system according to embodiment 92, furthercomprising a second shipping receptacle portion for receiving saidplurality of said paired tissue samples with data collections.

94. The portable transporter system according to embodiment 92, furthercomprising at least one fixation-inhibiting cooling device in thermalcommunication with said fixative in said holding chamber, wherein saidfixation-inhibiting cooling device is operable to reduce a temperatureof said fixative to inhibit fixation of the tissue sample contactingsaid fixative.

95. The portable transporter system according to embodiment 94, furthercomprising at least one cold transport box operable to receive saidfirst shipping receptacle portion, said plurality of said paired tissuesamples with data collections, and said at least one fixation-inhibitingcooling device.

96. The portable transporter system according to embodiment 76, whereinsaid cassette septum comprises at least one split.

97. The portable transporter system according to embodiment 96, whereinsaid at least one split of said cassette septum comprises at least twosplits.

98. The portable transporter system according to embodiment 97, whereinsaid at least two splits of said cassette septum comprises wherein saidat least two splits are perpendicular to one another.

99. The portable transporter system according to embodiment 97, whereinsaid at least two splits of said cassette septum comprises wherein saidat least two splits are of different length to one another.

100. The portable transporter system according to embodiment 96, whereinsaid at least one split of said cassette septum comprises at least onecross-shaped split.

101. The portable transporter system according to embodiment 76, whereinthe standard histology cassette holding the tissue sample is operable tomaintain the tissue sample submersed in fluid independent of theorientation of the transport container.

102. The portable transporter system according to embodiment 76, whereinsaid cassette clip comprises a clip feature operable to hold and retainthe standard histology cassette, where the standard histology cassetteis adapted to include a corresponding feature.

103. The portable transporter system according to embodiment 76, furthercomprising a carrier feature operable to slideably receive and retainthe standard histology cassette therein.

104. The portable transporter system according to embodiment 76, whereinsaid container lid comprises at least one side operable to hold anunused cassette upward to avoid contamination with said fixative priorto sample insertion.

105. The portable transporter system according to embodiment 76, whereinsaid container lid comprises at least one side operable to seal saidfixative fluid inside said holding chamber of said transport containerduring transport or handling and to avoid spillage and fumes.

106. The portable transporter system according to embodiment 76, whereinsaid container lid comprises at least one side operable to releasablycouple said container lid onto said holding chamber.

107. The portable transporter system according to embodiment 106,wherein said at least one side operable to releasably couple saidcontainer lid onto said holding chamber couples via matching screwthreads.

108. The portable transporter system according to embodiment 76, whereinthe standard histology cassette is held separately until the tissuesample is collected, and wherein said container lid is operable toreceive the standard histology cassette when the tissue sample is readyfor insertion into said holding chamber.

109. The portable transporter system according to embodiment 76, whereinsaid container lid comprises:

-   -   a multi-purpose lid wherein at least one side is operable to at        least one of:    -   hold an unused cassette upward to avoid contamination with said        fixative prior to sample insertion;    -   seal said fixative fluid inside said holding chamber of said        transport container during transport or handling and to avoid        spillage and fumes;    -   releasably couple said container lid onto said holding chamber;    -   releasably couple said container lid onto said holding chamber        couples via matching screw threads; or    -   receive the cassette when the tissue sample is ready for        insertion.

110. The portable transporter system according to embodiment 76, furthercomprising an electronic data logger coupled to said transport containervia a pod.

111. The portable transporter system according to embodiment 110,wherein said electronic data logger is in thermal communication withsaid transport container.

112. The portable transporter system according to embodiment 110,wherein said electronic data logger is operable to at least one of:

-   -   sense insertion of the tissue sample in said transport        container;    -   determine time of insertion of the tissue sample data;    -   store time of insertion of the tissue sample data in at least        one memory;    -   log time and date data of at least one of:        -   insertion in said transport container of the tissue sample            in the standard histology cassette, or        -   receipt of a start time data;    -   start a timer to track time data from at least one of:        -   an insertion time of the standard histology cassette with            the tissue sample in said transport container, or        -   receipt of a start timer data signal;    -   determine a time duration data of at least one of:        -   from said log time and date, or        -   from said start of said timer;    -   indicate a time duration data since at least one of:        -   said log time and date, or        -   said start of said timer;    -   indicate data via a display;    -   indicate data via a liquid crystal display (LCD);    -   indicate data via a light emitting diode (LED) display;    -   record a temperature data;    -   record a temperature and time data;    -   record at least one of a temperature or a time of crossing at        least one temperature threshold data;    -   record at least one of a temperature or time of temperature        excursion data against at least one preprogrammed threshold;    -   record at least one of a temperature or time of temperature        excursion against heating or freezing data;    -   record at least one temperature change and a time of said at        least one temperature change data;    -   indicate at least one recording;    -   indicate on at least one display said at least one record;    -   monitor temperature excursions data;    -   monitor at least one temperature change, and a time of said at        least one temperature change, and log said at least one        temperature change and said time data;    -   monitor data of at least one temperature change and time after        seal of said transport container, and indicate relative        performance of said data against at least one pre-programmed        threshold;    -   indicate on at least one display results data of said monitor;    -   store data on at least one memory;    -   store at least one of sensed data; recorded data; monitored        data; or calculated data on at least one memory;    -   store date on at least one memory, wherein said data comprises        at least one of:        -   a sample identifier;        -   an identifier;        -   a radio frequency identifier (RFID);        -   a bar code identifier;        -   a QR code identifier;        -   a location;        -   a sensed location;        -   a received location;        -   a global positioning system (GPS) location;        -   a location relative to a Wi-Fi access point;        -   a location relative to a wireless communication network;        -   a wired communication data;        -   a wireless communication data;        -   a direct connection;        -   a local connection;        -   a local area network(LAN);        -   a wide area network (WAN);        -   a remote connection;        -   a Bluetooth network data;        -   a near field communication (NFC) data;        -   a Zigbee protocol-compliant standard wireless communication            data;        -   a date and time of a data point;        -   a sensed data point;        -   a port connection;        -   a universal serial bus (USB) port connection;        -   a coupling to a communication device;        -   a coupling to a computing device;        -   a coupling to a portable device;        -   a coupling to a wireless device;        -   a coupling to a personal computer device;        -   a coupling to a smartphone device;        -   a coupling to a tablet device;        -   a coupling to a mobile phone device; or        -   a coupling to a telephony device.

113. The portable transporter system according to embodiment 76, furthercomprising a carrier unit integrated with a plurality of said containerlids.

114. The portable transporter system according to embodiment 113,wherein said integrated carrier unit comprises wherein said plurality ofsaid container lids comprises at least four container lids.

115. The portable transporter system according to embodiment 113,further comprising a protective sheath operable to protect the standardhistology cassette against contamination or dust.

116. The portable transporter system according to embodiment 113,wherein said carrier unit comprises a plurality of holding chambersshipped with fixative in said plurality of holding chambers, each havingone of said container lids.

117. The portable transporter system according to embodiment 113,wherein said container clip does not require the standard histologycassette to be removed from said container lid.

118. The portable transporter system according to embodiment 113,wherein said container lid is reversible and can be screwed onto saidtransport container immersing the tissue sample in said fixative.

119. The portable transporter system according to embodiment 113,wherein it is possible at a glance to see which of said plurality ofholding chambers have samples inserted.

120. The portable transporter system according to embodiment 113,wherein said fixative comprises formalin.

121. The portable transporter system according to embodiment 113,wherein each said container lid is tall enough to receive application ofat least one label, code, barcode, or identifier.

122. The portable transporter system according to embodiment 113,wherein each said transport container cannot be inadvertently leftoutside of a fixation-inhibiting cooling device.

123. The portable transporter system according to embodiment 113,wherein said cassette clip holds the standard histology cassette to easereading of at least one code on the standard histology cassette.

124. The portable transporter system according to embodiment 76, whereinsaid fixative is prefilled into said transport container, and isrefrigerated to be ready for use.

125. The portable transporter system according to embodiment 124,wherein the tissue sample, when surgically removed is marked with apatient identifier.

126. The portable transporter system according to embodiment 125,wherein the patient identifier comprises at least one of:

-   -   a barcode;    -   a wristband; or    -   a radio frequency identifier (RFID).

127. The portable transporter system according to embodiment 126,wherein the tissue sample is inserted into the standard histologycassette, and the standard histology cassette is placed in saidtransport container, wherein the standard histology cassette has aunique identifier.

128. The portable transporter system according to embodiment 127,wherein said transport container is transferred into a pod, wherein saidpod holds a data logger in close proximity, and wherein said data loggerlogs at least one start time and start temperature reading.

129. The portable transporter system according to embodiment 128,wherein said pod is inserted into a fixation-inhibiting cooling device.

130. The portable transporter system according to embodiment 129,wherein said fixation-inhibiting cooling device ensures the tissuesample is kept at 4° C. during transport.

131. The portable transporter system according to embodiment 129,wherein said data logger logs temperatures continuously during transportto a laboratory.

132. The portable transporter system according to embodiment 129,wherein said data logger logs at least one of a temperature or a time atleast one of:

-   -   continuously;    -   periodically;    -   at least one threshold;    -   at least one temperature threshold;    -   at a time interval;    -   upon a temperature change;    -   according to sensed data; or    -   after a time duration.

133. The portable transporter system according to embodiment 129,wherein upon arrival at the laboratory, time and temperature data isread from said data logger of said pod of said transport container andverifies integrity of the tissue sample.

134. The portable transporter system according to embodiment 133,wherein the tissue sample is stored in cold storage if additional coldstorage time is determined to be required.

135. The portable transporter system according to embodiment 133,wherein the tissue sample is reviewed and at least one of transferred orkept in a final code identified standard histology cassette.

136. The portable transporter system according to embodiment 135,wherein the tissue sample is processed using a standard heated fixativeprocessor, and with subsequent processing steps.

137. The portable transporter system according to embodiment 136,wherein the standard heated fixative processor processing the tissuesample for 2 hours at 45° C. temperature.

138. The portable transporter system according to embodiment 136,wherein said subsequent processing steps comprises at least one of analcohol processing step, a xylene processing step, or a paraffinprocessing step.

139. The portable transporter system according to embodiment 136,wherein the tissue sample is embedded, sectioned, and stained.

140. The portable transporter system according to embodiment 139,wherein the tissue sample is stained with various markers or probes.

141. The portable transporter system according to embodiment 139,wherein the stain results are made available for at least one of:

-   -   review by a pathologist,    -   receipt of at least one score by a pathologist; or    -   report results for the tissue sample to at least one clinical        database.

142. The portable transporter system according to embodiment 76, furthercomprising wherein electronic data is collected.

143. The portable transporter system according to embodiment 142,wherein said electronic data is collected by at least one of:

-   -   an electronic pen;    -   a unique identifier;    -   a tablet device;    -   a wireless identifier;    -   a Wi-Fi media access control (MAC) address;    -   a static computing device;    -   a mobile computing device;    -   a barcode scanner;    -   an external data temperature logger;    -   a portable fixation-inhibiting cooling device comprising        intelligence;    -   a portable fixation-inhibiting cooling device comprising an        electronic datalogger for collecting temperature and time data;

144. The portable transporter system according to embodiment 76, whereina fixation-inhibiting cooling element is pre-charged over night for nextday surgery.

145. The portable transporter system according to embodiment 76, whereinsaid transport container is kept pre-cooled in a refrigerator.

146. The portable transporter system according to embodiment 76, whereina fixation-inhibiting cooling device is configured prior to surgery, istransported to surgery location, and keeps reagents cold.

147. The portable transporter system according to embodiment 76, whereintransportation of said portable transporter system is performed at anend of a surgery day.

148. The portable transporter system according to embodiment 76, whereina fixation processor is operable to at least one of:

-   -   collect transport data;    -   perform final fixation steps required; or    -   provide a comprehensive fixation quality report for each of the        tissue samples.

149. The portable transporter system according to embodiment 142,wherein said electronic data is collected by an electronic pen, andcollection comprises processing of data of the tissue sample comprisingat least one of:

-   -   establish chain-of-custody during the tissue sample collection,        integrate data comprising at least one of: patient-ID, location        of surgery, day/time, name of surgeon, or location of removal of        the tissue sample;    -   associate with a designated of said transport container and        cassette identifier (ID);    -   record insertion time of the tissue sample into the transport        container;        -   log transport conditions comprising at least one temperature            or time profile;    -   document a total time of cold diffusion after which the sample        can be safely removed and submitted to a heated fixative step        for crosslinking;    -   duplicate capture of handwritten Lab Requisition Form by        electronic data capture and associate data to a specific        specimen of the tissue sample or the standard histology cassette        via identifier comprising at least one of a barcode or        radio-frequency identifier (RFID);    -   verify a minimum fixation condition; or    -   enable data capture for a clinical trial.

150. The portable transporter system according to embodiment 149,wherein said electronic data is collected by an electronic pen (ePen)comprising at least one of:

-   -   use ePen to fill out lab requisition form;    -   collect electronic data;    -   collect identifier information from patient, said transport        container, the standard histology cassette;    -   transfer electronic data to a computing device for verification        or correction;    -   upload verified electronic data via communications network if        possible;    -   if upload not possible, ePen travels with the tissue sample in        fixation-inhibited cooling device;    -   receive at accessioning stage in histology lab for display and        report to the LIS;    -   optionally enter any corrections, if required; or    -   provide readout of datalogger, or integrate into data uploaded        to the LIS.

151. The portable transporter system according to embodiment 142,wherein said electronic data is collected by a unique identifier.

152. The portable transporter system according to embodiment 151,wherein said unique identifier comprises at least one of:

-   -   an Anoto code printed onto a label of a container;    -   a barcode on the cassette could be a vantage compatible        preprinted code being inserted into said container;    -   a barcode on the cassette identifiable by a barcode reader when        said container arrives at accessioning stage, wherein said        barcode reader is tied to patient-ID associated to a code of        said container; or    -   a unique label with Anoto pattern on said container, wherein the        container is picked randomly, pen ties code electronically to        patient-ID, and handwritten link and an identifier to be copied        from a label of said container onto paper.

153. The portable transporter system according to embodiment 142,wherein said electronic data is collected by a tablet device coupled tothe portable transporter system by at least one of wired or wirelesscommunication, and collection comprises processing of data of the tissuesample comprising at least one of:

-   -   establish chain-of-custody during the tissue sample collection,        integrate data comprising at least one of: patient-ID, location        of surgery, day/time, name of surgeon, or location of removal of        the tissue sample;    -   associate with a designated of said transport container and        cassette identifier (ID);    -   record insertion time of the tissue sample into the transport        container;        -   log transport conditions comprising at least one temperature            or time profile;    -   document a total time of cold diffusion after which the sample        can be safely removed and submitted to a heated fixative step        for crosslinking;    -   duplicate capture of handwritten Lab Requisition Form by        electronic data capture and associate data to a specific        specimen of the tissue sample or the standard histology cassette        via identifier comprising at least one of: a barcode; or        radio-frequency identifier (RFID);    -   data capture via at least one of: camera; RFID; or near field        communication

(NFC) tag via communication interface;

-   -   capture of image of surgical site;    -   verify a minimum fixation condition; or    -   enable data capture for a clinical trial.

154. The portable transporter system according to embodiment 153,wherein said electronic data is collected by said tablet comprising atleast one of:

-   -   capture pertinent information about patient and study on the        tablet;    -   capture duplicate entry of lab requisition form on the tablet;    -   optionally correct any data entry errors on screen, if needed;    -   optionally upload verified electronic data via wireless        communication to LIS;    -   provide tablet accompanying the tissue sample in same cooling        and shipping box;    -   receive at the accessioning stage in histology lab the tablet        for display or report to the LIS; or    -   provide readout of datalogger, or integrate into data uploaded        to the LIS.

155. The portable transporter system according to embodiment 142,wherein said electronic data is collected by a wireless identifier.

156. The portable transporter system according to embodiment 155,wherein said electronic data is collected by said wireless identifiercomprising a wireless fidelity (Wi-Fi) media access control (MAC)address.

157. The portable transporter system according to embodiment 156,wherein said electronic data is collected by said Wi-Fi MAC address andwherein a device having said Wi-Fi MAC address comprises at least oneof:

-   -   a datalogger and positional time stamper operable to track the        tissue sample;    -   any time a Wi-Fi router or device with said MAC address        communicate, identity of both said router and said device are        logged; or    -   since time of contact is known and location of a Wi-Fi router is        known, a location of a device with said MAC address is recorded        over the journey of the MAC device via a network of free-access        Wi-Fi routers.

158. The portable transporter system according to embodiment 142,wherein said electronic data is collected by a static computing device,and collection comprises processing of data of the tissue samplecomprising at least one of:

-   -   establish chain-of-custody during the tissue sample collection,        integrate data comprising at least one of: patient-ID, location        of surgery, day/time, name of surgeon, or location of removal of        the tissue sample;    -   associate with a designated of said transport container and        cassette identifier (ID);    -   record insertion time of the tissue sample into the transport        container;        -   log transport conditions comprising at least one temperature            or time profile;    -   document a total time of cold diffusion after which the sample        can be safely removed and submitted to a heated fixative step        for crosslinking;    -   duplicate capture of handwritten Lab Requisition Form by        electronic data capture and associate data to a specific        specimen of the tissue sample or the standard histology cassette        via identifier comprising at least one of: a barcode; or        radio-frequency identifier (RFID);    -   data capture via at least one of: camera; RFID; or near field        communication (NFC) tag via communication interface;    -   capture of image of surgical site;    -   verify a minimum fixation condition; or    -   enable data capture for a clinical trial.

159. The portable transporter system according to embodiment 158,wherein said electronic data is collected by said static computingdevice comprises at least one of:

-   -   capture pertinent information about patient and study;    -   capture duplicate entry of lab requisition form;    -   optionally correct any data entry errors on screen, if needed;    -   optionally upload verified electronic data via communication        network to LIS;    -   if LIS connectivity is prohibited, optionally program data        logger to retain additional information on computer locally;    -   receive at the accessioning stage in histology lab identifier        information to reference the tissue sample via LIS query for        display or report to the LIS; or    -   provide readout of datalogger, or integrate into data uploaded        to the LIS.

160. The portable transporter system according to embodiment 142,wherein said electronic data is collected by a travelling coolingshipping box with intelligence, and collection comprises processing ofdata of the tissue sample comprising at least one of:

-   -   establish chain-of-custody during the tissue sample collection,        integrate data comprising at least one of: patient-ID, location        of surgery, day/time, name of surgeon, or location of removal of        the tissue sample;    -   associate with a designated of said transport container and        cassette identifier (ID);    -   record insertion time of the tissue sample into the transport        container;        -   log transport conditions comprising at least one temperature            or time profile;    -   document a total time of cold diffusion after which the sample        can be safely removed and submitted to a heated fixative step        for crosslinking;    -   duplicate capture of handwritten Lab Requisition Form by        electronic data capture and associate data to a specific        specimen of the tissue sample or the standard histology cassette        via identifier comprising at least one of: a barcode; or        radio-frequency identifier (RFID);    -   data capture via at least one of: camera; RFID; or near field        communication (NFC) tag via communication interface;    -   capture of image of surgical site;    -   verify a minimum fixation condition; or    -   enable data capture for a clinical trial.

161. The portable transporter system according to embodiment 160,wherein said electronic data is collected by said travelling coolingshipping box with said intelligence, comprising at least one of:

-   -   capture pertinent information about patient and study;    -   capture duplicate entry of lab requisition form;    -   optionally interface computer with a host computer in said        travelling cooling shipping box to store data;    -   optionally monitor loading or unloading of the tissue sample by        the host computer in said travelling cooling shipping box, and        log time and temperature;    -   optionally correct any data entry errors on screen, if needed;        or    -   at the accessioning stage in histology lab, the host computer        provides electronic data collected.

162. The portable transporter system according to embodiment 142,wherein said transport of the tissue sample comprises a cooling boxconfigured to at least one of:

-   -   pre-charge over night for a next day surgery;    -   pre-cool in fridge said container;    -   prior to surgery transport to surgery location and maintain in        cold temperature reagents; or    -   initiate shipping at end of surgery day.

163. The portable transporter system according to embodiment 162,wherein said cooling box is maintained at about 4° C.

164. A portable transporter system adapted to carry a tissue samplecontacting a fixative, comprising:

-   -   a transport container comprising:    -   a holding chamber; and    -   a fixative; and    -   a container lid comprising:    -   a cassette holder operable to be coupled to the standard        histology cassette and comprising a first seal; and    -   a cassette receiver operable to be releasably coupled to said        transport container and comprising a second seal,    -   wherein said cassette holder and said cassette receiver are        operable to be attachably coupled to one another upon insertion        of the standard histology cassette coupled to said cassette        holder into said cassette receiver.

165. The portable transporter system according to embodiment 164,wherein said cassette holder and said cassette receiver are operable tobe attachably coupled to one another via at least one male snap clip andat least one female snap clip receiver.

166. The portable transporter system according to embodiment 165,wherein said at least one male snap clip is on a bottom portion of saidcassette holder, and said at least one female snap clip receiver is onan upper portion of said cassette receiver.

167. The portable transporter system according to embodiment 165,wherein said at least one male snap clip comprises a plurality of saidat least one male snap clips.

168. The portable transporter system according to embodiment 167,wherein said plurality of said male snap clips comprises at least foursnap clips.

169. The portable transporter system according to embodiment 164,wherein said first and second seals comprise solvent compatible o-rings.

170. The portable transporter system according to embodiment 164,wherein said fixative has a fixative volume comprising a holding chambervolume of said holding chamber less a cassette volume of a standardhistology cassette.

171. A portable transporter system adapted to carry a tissue contactinga fixative, comprising:

-   -   a transport container comprising:    -   a holding chamber; and    -   a container lid operable to be releasably coupled to said        transport container, said container lid comprising:    -   a built-in septum comprising a molded boss comprising an        external thread; and    -   a standard sealing screw-top comprising an inner thread matching        said external thread of said molded boss, said screw-top        operable to cover said built-in septum.

172. The portable transporter system according to embodiment 171,further comprising a syringe comprising a pre-filled amount of fixative,and operable to be refrigerated until used to fill said holding chamberwith said fixative, via said built-in septum, after the tissue sample isplaced inside said holding chamber.

173. The portable transporter system according to embodiment 172,wherein said fixative comprises formalin.

174. The portable transporter system according to embodiment 171,further comprising a cassette clip attachably couplable to receive, holdand retain the standard histology cassette.

175. A transporter system for protecting biomarkers in a tissue sample,comprising:

-   -   a specimen container configured to retain or hold at least one        tissue sample in a reagent, wherein the reagent is chilled to        from about 0° C. to about 15° C. prior to transportation to a        laboratory;    -   a carrier assembly configured to retain or hold the specimen        container; and    -   a monitoring system that comprises at least one sensor        configured to obtain, store or transmit one or both of time and        temperature information about the tissue sample and/or reagent        in the specimen container when the specimen container is        positioned in the carrier assembly and transported to a        laboratory, wherein the monitoring system further comprises a        data logging device.

176. The transporter system of embodiment 175 wherein the reagent is atissue fixation reagent.

177. The transporter system of embodiment 175 further comprising atissue sample cassette for retaining one or more tissue samples thereinand wherein the specimen container is configured to retain and hold thetissue sample cassette in the reagent.

178. The transporter system of embodiment 1751, wherein the monitoringsystem measures the temperature of the reagent and/or tissue sample inthe specimen container over a period of time.

179. The transport system of embodiment 175, further comprising atransport container for transporting the specimen container, carrierassembly and monitoring system to a destination.

180. The transporter system of embodiment 175, wherein the monitoringsystems comprises one or more sensors selected from the group consistingof temperature, elapsed time, transporter temperature, ambient light,infrared proximity, accelerometer, acoustic, and capacitive sensors.

181. The transporter system of embodiment 175, wherein the monitoringsystem is physically coupled to the carrier assembly.

182. The transporter system of embodiment 175, wherein the monitoringsystem is electronically coupled to the carrier assembly.

183. The transporter system of embodiment 175, wherein the monitoringsystem comprises at least one sensor, wherein the sensor is atemperature sensor positioned in the specimen container and comprises atransmitter that outputs a temperature signal to a receiver located inthe carrier assembly holding that specimen container.

184. The transporter system of embodiment 175, wherein the monitoringsystem either stores temperature measurements indicative of thetemperature of the reagent and/or the tissue sample or transmits suchinformation to an external receiver wherein the receiver comprises amemory device.

185. The transporter system of embodiment 4, wherein the specimencontainer comprises a temperature sensor wherein said sensor transmitsthe temperature of the reagent contained therein to the monitoringsystem.

186. The transport system of embodiment 175, wherein the specimencontainer further comprises a radio frequency identification (RFID) tagand the monitoring system further comprises a RFID reader.

187. The transport system of embodiment 175, further comprising atransport container for transporting the specimen container, carrierassembly and monitoring system wherein the transport container comprisesan ice bath.

188. The transport system of embodiment 175, further comprising atransport container for transporting the specimen container, carrierassembly and monitoring system, wherein the transport container isconfigured to maintain the temperature of a specimen container thereinto from about 0° C. to about 15° C. for up to 14 days.

189. The transporter system of embodiment 175, further comprising aplurality of carrier assemblies and a plurality of specimen containerswherein each of the plurality of carrier assemblies holds a separate oneof the plurality of specimen containers.

190. The transporter system of embodiment 175 further comprises acooling device configured to reduce or maintain an average temperatureof reagent in the specimen container to or at a temperature equal to orlower than about 15° C.

191. An apparatus for transporting tissue samples, comprising:

-   -   a plurality of specimen containers holding reagents;    -   a plurality of carrier assemblies, each assembly comprising a        specimen container holder and a sensor assembly positioned to        monitor the temperature of the specimen container or the reagent        held in each specimen container and time the container is held        in the holder, wherein the sensor assembly comprises a        temperature sensor and a time sensor;    -   a transport assembly for carrying the multiple carrier        assemblies; and    -   a transporter for carrying at least one transport assembly.

192. The apparatus of embodiment 175, wherein the sensor assembly ofeach of the plurality of carrier assemblies comprises a temperaturesensor positioned within the specimen container, wherein the temperaturesensor includes a transmitter that outputs a temperature signal to areceiver located in the carrier assembly holding that specimencontainer.

193. The apparatus of embodiment 192, further comprising a computingdevice communicatively coupled to the transmitters and receivers,wherein the computing device is configured to evaluate sound speeds inthe tissue sample by evaluating time of flight changes of acoustic wavesfrom one or more of the transmitters.

194. The apparatus of embodiment 192, further comprising a computingdevice configured to compensate for changes in the time of flight ofacoustic waves in a fixative or sample associated with environmentalchanges.

195. The apparatus of embodiment 192, further comprising the computingdevice containing instructions, when executed, to cause the computingdevice to compensate for temperature changes of one or more of thereagent or sample.

196. A transporter system for carrying a tissue sample, comprising:

-   -   a specimen container configured to hold a tissue sample in a        fixative;    -   a carrier assembly configured to retain or hold a specimen        container;    -   a monitoring system that obtains and stores time and temperature        information about the tissue sample in the specimen container        when the specimen container is positioned in the carrier        assembly; and    -   a transport container configured to hold the carrier assembly        holding the specimen container and monitoring system, wherein        the transport container is further configured to maintain the        temperature of the internal holding chamber at from about 0° C.        to about 20° C. for at least 1 hour.

197. The transporter system of embodiment 196, wherein the transportcontainer is configured to maintain the temperature of the internalholding chamber at from about 0° C. to about 15° C. for at least 24hours.

198. The transporter system of embodiment 196, wherein the transportcontainer is configured to maintain the temperature of the internalholding chamber at from about 0° C. to about 15° C. for up to 14 days.

199. The transporter system of embodiment 196, wherein the carrierassembly is configured to hold a plurality of specimen containers eachholding a respective tissue sample, wherein the monitoring systemincludes a plurality of temperature sensors each measuring a temperatureof the tissue sample in one of the specimen containers.

200. The transporter system of embodiment 196, wherein the monitoringsystem measures the temperature of the fixative and/or tissue sampleover a period of time.

201. The transporter system of embodiment 196, wherein the monitoringsystem includes a temperature sensor carried by the specimen containerand positioned to measure a temperature of the fixative.

202. The transporter system of embodiment 196, wherein the monitoringsystem stores temperature measurements indicative of the temperature ofthe fixative and/or the tissue sample.

203. The transport system of embodiment 196, wherein the monitoringsystem includes a reader physically coupled to the carrier assembly anda radio-frequency identification tag coupled to the specimen container.

204. The transporter system of embodiment 196, wherein the transportcontainer comprises a cooling device configured to reduce an averagetemperature of fixative in the specimen container from room temperatureto a temperature equal to or lower than about 15° C.

205. The transporter system of embodiment 196, wherein the monitoringsystem further comprises a timer.

206. The transporter system of embodiment 196, wherein the specimencontainer includes a cover and a main body, wherein the cover includes aholder configured to hold the tissue sample in contact with thefixative.

207. A method for processing a tissue sample, comprising:

-   -   contacting the tissue sample with a fixative chilled to from        about 0° C. to about 15° C., wherein the fixative is contained        in a specimen receiving container;    -   placing a specimen container in a holding well of a carrier        assembly further comprising at least one data logging device;    -   detecting and storing time and temperature information        associated with the tissue sample in the data logger of the        carrier assembly;    -   placing the carrier assembly in a transport assembly; and    -   transporting the transport assembly to a destination.

208. The method of embodiment 207, wherein detecting the time andtemperature information associated with the tissue sample includes:

-   -   measuring a temperature of the fixative and/or the tissue        sample, and    -   measuring a contact period of time in which the fixative        contacts the tissue sample.

The method of embodiment 33, wherein detecting the time and temperatureinformation associated with the tissue sample includes obtainingtemperature measurements and storing the temperature measurements.

The various embodiments and features described above can be combined toprovide further embodiments. These and other changes can be made to theembodiments in light of the above-detailed description. In general, inthe following claims, the terms used should not be construed to limitthe claims to the specific embodiments disclosed in the specificationand the claims, but should be construed to include all possibleembodiments along with the full scope of equivalents to which suchclaims are entitled. Accordingly, the claims are not limited by thedisclosure.

1. A tissue sample transport and storage assembly comprising: (a) asample container assembly comprising: (a1) walls defining a first cavityfor holding a volume of a formalin-based fixative solution; (a2) ahistology cassette sized for holding the tissue sample and adapted forinsertion into the cavity of the sample container, such that the tissuesample is completely immersed into the volume of the formalin-basedfixative solution; and (a3) a seal adapted to create aformalin-impervious barrier enclosing the cavity; (b) atemperature-responsive element configured to track and record thetemperature, of a formalin-based fixative solution disposed in thesample container, through the walls and/or seal; and (c) a case assemblycomprising a second cavity defined by a material comprising an insulatorand/or a cooling element, wherein the cavity is sized to accept thesample container and the temperature-responsive element in anarrangement that permits the temperature-responsive element to track andrecord the temperature of the formalin-based fixative solution in thesample container, and wherein the insulator and/or a cooling element iscapable of maintaining the volume of the formalin-based fixativesolution at a temperature in the range of 0° C. to 7° C. for at least 2hours under shipping conditions when the tissue sample transport andstorage assembly is assembled.
 2. The tissue sample transport andstorage assembly of claim 1, wherein the seal is a container lidcomprising a cassette clip attachably couplable to receive, hold andretain the histology cassette.
 3. The tissue sample transport andstorage assembly of claim 2, wherein said cassette clip is removablycouplable to said container lid.
 4. The tissue sample transport andstorage assembly of claim 2, wherein said cassette clip is configured tobe screwed into at least one side of said container lid.
 5. The tissuesample transport and storage assembly of claim 2, wherein the cassetteclip is configured to be clipped to the histology cassette.
 6. Thetissue sample transport and storage assembly of claim 2, wherein saidcassette clip comprises: a first end operable to be coupled to at leastone side of said container lid; and a second end operable to be coupledto the standard histology cassette.
 7. The tissue sample transport andstorage assembly of claim 1, further comprising a cassette coverremovably coupled to said container lid.
 8. The tissue sample transportand storage assembly of claim 7, wherein said cassette cover comprises asterile seal between said cassette cover and said container lid.
 9. Thetissue sample transport and storage assembly of claim 1, wherein thesample container assembly further comprises a cassette septum, whereinthe cassette septum is removable with said container lid, said cassetteclip, and the standard histology cassette.
 10. The tissue sampletransport and storage assembly of claim 9, wherein said cassette septumcomprises a breakable membrane.
 11. The tissue sample transport andstorage assembly of claim 10, wherein said container lid furthercomprises an arm operable to break said breakable membrane uponinsertion of the cassette into said holding chamber of said transportcontainer.
 12. The tissue sample transport and storage assembly of claim2, wherein said container lid further comprises a cylindrical portioncomprising an internal screw thread portion adapted to couple with anexternal screw thread portion of said transport container, operable toisolate the sample, air and said fixative from a surroundingenvironment.
 13. The tissue sample transport and storage assembly ofclaim 1, wherein the material defining the second cavity comprises acooling element.
 14. The tissue sample transport and storage assembly ofclaim 13, wherein the cooling element comprises at least one elementselected from the group consisting of: a first substance and a secondsubstance that mix to produce an endothermic reaction in response to thefixation-inhibiting cooling device changing from a non-cooling mode to acooling mode; a desiccant and a substance that evaporates to cool thefixative; and a refrigeration unit or a thermoelectric cooler unit. 15.The tissue sample transport and storage assembly of claim 13, whereinthe cooling element comprises a cooling material having a freezing pointin the range of 0° C. to 7° C.
 16. The tissue sample transport andstorage assembly of claim 15, wherein the volume of the cooling materialis selected such that the time for the volume of coolant to transitionfrom completely frozen to completely melted is at least two hours whenstored at 20° C. without pressurization.
 17. The tissue sample transportand storage assembly of claim 1, wherein said temperature responsiveelement comprises an absorbent material and a dye, wherein the dye candiffuse through the absorbent material when at a first temperature rangeand to not diffuse through the dye when at a second temperature range,wherein the extent of diffusion of the dye through the absorbentmaterial correlates with the amount of time that the formalin-basedfixative solution has been at a temperature in the first temperaturerange.
 18. The tissue sample transport and storage assembly of claim 17,wherein the first temperature range is from 0° C. to 7° C., and thesecond temperature range is a temperature above 7° C.
 19. The tissuesample transport and storage assembly of claim 17, wherein the dye isdisposed in a matrix having a melting point at a temperature above 7°C., wherein the first temperature range is a temperature at or above themelting point, wherein the second temperature range is a temperaturebelow the melting point.
 20. The tissue sample transport and storageassembly of claim 1, wherein said temperature responsive elementcomprises: a first absorbent material and a first dye, wherein the firstdye is engineered to diffuse through the absorbent material when attemperature range is from 0° C. to 7° C. and to not diffuse through theabsorbent material when at a temperature above 7° C., and wherein theextent of diffusion of the first dye through the first absorbentmaterial correlates with the amount of time that the formalin-basedfixative solution has been at a temperature in the temperature rangefrom 0° C. to 7° C.; a second absorbent material and a second dye,wherein the second dye is disposed in a matrix having a melting point ata temperature above 7° C., wherein the second dye begins diffusingthrough the second absorbent material when the matrix begins melting,and wherein the extent of diffusion of the second dye through the secondabsorbent material correlates with the amount of time that theformalin-based fixative solution has been at a temperature above themelting point.
 21. The tissue sample transport and storage assembly ofclaim 20, wherein the absorbent material is attached to or adapted to beattached to the walls or the seal of the sample container assemblywithout being placed within the cavity defined by the walls.
 22. Thetissue sample transport and storage assembly of claim 1, furthercomprising a data logger.
 23. The tissue sample transport and storageassembly of claim 22, wherein the temperature responsive elementcomprises a temperature sensor adapted to communicate the temperature ofthe formalin-based fixative solution to the data logger.
 24. The tissuesample transport and storage assembly of claim 23, wherein thetemperature sensor is an infrared temperature sensor.
 25. The tissuesample transport and storage assembly of claim 22, wherein the datalogger further comprises a timer.
 26. The tissue sample transport andstorage assembly of claim 22, wherein the data logger is operable to atleast one of: sense insertion of the tissue sample in said transportcontainer; determine time of insertion of the tissue sample data; storetime of insertion of the tissue sample data in at least one memory; logtime and date data of at least one of: insertion in said transportcontainer of the tissue sample in the standard histology cassette, orreceipt of a start time data; start a timer to track time data from atleast one of: an insertion time of the standard histology cassette withthe tissue sample in said transport container, or receipt of a starttimer data signal; determine a time duration data of at least one of:from said log time and date, or from said start of said timer; indicatea time duration data since at least one of: said log time and date, orsaid start of said timer; indicate data via a display; indicate data viaa liquid crystal display (LCD); indicate data via a light emitting diode(LED) display; record a temperature data; record a temperature and timedata; record at least one of a temperature or a time of crossing atleast one temperature threshold data; record at least one of atemperature or time of temperature excursion data against at least onepreprogrammed threshold; record at least one of a temperature or time oftemperature excursion against heating or freezing data; record at leastone temperature change and a time of said at least one temperaturechange data; indicate at least one recording; indicate on at least onedisplay said at least one record; monitor temperature excursions data;monitor at least one temperature change, and a time of said at least onetemperature change, and log said at least one temperature change andsaid time data; monitor data of at least one temperature change and timeafter seal of said transport container, and indicate relativeperformance of said data against at least one pre-programmed threshold;indicate on at least one display results data of said monitor; storedata on at least one memory; store at least one of sensed data; recordeddata; monitored data; or calculated data on at least one memory; storedata on at least one memory, wherein said data comprises at least oneof: a sample identifier; an identifier; a radio frequency identifier(RFID); a bar code identifier; a QR code identifier; a location; asensed location; a received location; a global positioning system (GPS)location; a location relative to a Wi-Fi access point; a locationrelative to a wireless communication network; a wired communicationdata; a wireless communication data; a direct connection; a localconnection; a local area network(LAN); a wide area network (WAN); aremote connection; a Bluetooth network data; a near field communication(NFC) data; a Zigbee protocol-compliant standard wireless communicationdata; a date and time of a data point; a sensed data point; a portconnection; a universal serial bus (USB) port connection; a coupling toa communication device; a coupling to a computing device; a coupling toa portable device; a coupling to a wireless device; a coupling to apersonal computer device; a coupling to a smartphone device; a couplingto a tablet device; a coupling to a mobile phone device; or a couplingto a telephony device.
 27. A sample transport device comprising a samplecontainer, a cooling block, a data logger, and an insulated case,wherein: (a) said sample container is sized to contain a tissue sampleimmersed in a volume of a formalin-based fixative solution and isinserted into a cavity in the cooling block; (b) said cooling blockcontains a volume of a coolant and is inserted in a cavity in the datalogger; (c) said data logger comprises: (c1) a temperature sensorconfigured to monitor the temperature of the coolant and/or the fixativesolution; and (c2) a timer configured to measure an amount of timeselected from the group consisting of: (c1a) an amount of time that thecooling block has been inserted in the data logger; (c1b) an amount oftime that the temperature of the coolant and/or the fixative solutionhas been within a particular temperature range; and (d) said insulatedblock comprises a cavity in which the data logger is inserted.
 28. Thedevice of claim 27, wherein the coolant is a phase change materialhaving a melting temperature within the range of 0° C. to 7° C.
 29. Amethod of fixing an unfixed tissue sample comprising: (a) immersing theunfixed tissue sample in a volume of a formalin-based fixative solutionchilled to a temperature in the range of 0° C. to 7° C. disposed in atissue sample transport and storage assembly according to claim 1; (b)storing and/or shipping the tissue sample transport and storage assemblyor system over a period of time, wherein: (b1) the insulator and/orcooling element maintains the volume of the formalin-based fixativesolution at a temperature in the range of 0° C. to 7° C. at least untilthe formalin-based fixative solution has diffused throughout the unfixedtissue sample; and (b2) the temperature-responsive element tracks andrecords the temperature of the volume of formalin-based fixativesolution during storage and shipment; and (c) after the formalin-basedfixative solution has diffused throughout the unfixed tissue sample,exposing the tissue sample to a formalin-based fixative solution havinga temperature in the range of from 20° C. to 55° C. for a sufficientperiod of time to induce fixation of the tissue sample.
 30. The methodof claim 29, wherein the insulator and/or cooling element allows thetemperature of the volume of the formalin-based fixative solution torise above 7° C. after the formalin-based fixative solution has diffusedthroughout the unfixed tissue sample.
 31. The method of claim 30,wherein the cooling element comprises a coolant having a melting pointin the range of 0° C. to 7° C., wherein the volume of coolant isselected such that the time for the volume of coolant to transition fromcompletely frozen to completely melted is at least two hours when storedat 20° C. without pressurization.